China Wuhan Desheng Biochemical Technology Co., Ltd Company News

Tris (trihydroxymethylaminomethane) is a weak base with a pKa of 8.1 at room temperature of 25℃ and an effective buffer range of pH 7.0 ~ 9.2.The pH of aqueous solution of Tris alkali is about 10.5. Hydrochloric acid is generally added to adjust the pH value to the desired value, so that the buffer of this pH value can be obtained.Since Tris buffer is a weakly alkaline solution, DNA will be deprotonated in such a solution, thus improving its solubility.If the pH-adjusted acid solution is replaced with acetic acid, a "TAE buffer" (Tris/Acetate/EDTA) is obtained, while a "TBE buffer" (Tris/Borate/EDTA) is obtained by replacing it with boric acid.These two buffers are commonly used in nucleic acid electrophoresis experiments.TAE, TBE, etc. prepared by Tris are the most commonly used reagents for DNA electrophoresis, and TE (pH 8.0) is mainly used to dissolve DNA.(TE is a combination of Tris and EDTA.)1MTris-HCl6.8 and 1.5MTris-HCl8.8 are the most commonly used reagents for SDS-PAGE.   TRIS reagent   Among the conventional operations of genetic engineering, agarose gel electrophoresis is the most widely used.Agarose gel electrophoresis is a conventional method for separating, identifying and purifying DNA fragments.It usually uses a horizontal electrophoresis device to electrophoresis under an electric field with constant intensity and direction.DNA molecules are negatively charged in gel buffers (generally alkaline) and migrate from negative to positive electrodes in an electric field.The rate of DNA molecule migration is dependent on the size and conformation of the molecule.The influence of electric field strength and direction, base composition, temperature and embedded dyes, etc.     Operation process: ① TE buffer: (10 mmol/L Tris-HCl, 1 mmol/L EDTA) Electrophoresis buffer (50XTAE)   ② Electrophoresis buffer (50XTAE): Tris 242g, glacial acetic acid 57.1ml, EDTA (0.5mol/L pH 8.0) 100ml Dilute 50 times with distilled water when used.   ③ Sample buffer (6X): 0.25% bromophenol blue, 0.25% xylene blue, 40% (W/V) sucrose   ④ STET buffer (pH 8.0) (8% sucrose, 0.5% Triton, 50 mmol/L EDTA, 10 mmol/L Tris) 1) Measure 100 ml of TAE electrophoresis solution, add 0.7 g of agarose, mix well, place in microwave oven, heat for 3 minutes, fully dissolve agarose. 2) Close the clean and dry electrophoretic plate with sterilized rubber and seal the edge with a small amount of agarose solution.Place the comb and adjust the distance between the bottom edge of the comb and the electrophoresis plate, generally 1-2 mm is appropriate. 3) When the dissolved agarose solution is cooled to about 50C, add 5 M L of ethidium bromide, and the final concentration of ethidium bromide is 1.0 m g/m L. After mixing, pour the agarose solution into the electrophoresis plate and keep it still without moving. 4) After the gel has completely solidified (30-45 minutes at room temperature), gently remove the comb, remove the tape, and place the gel in the electrophoresis pad. 5) In the electrophoresis process, electrophoresis solution (1'TAE) was added to cover the agarose gel surface with about 1-2 mm electrophoresis solution.

3-(cyclohexylamine)-1-propanesulfonic acid, also called CAPS, is a kind of biological buffer, which is commonly used in biochemical diagnostic kit, DNA/RNA extraction kit and PCR diagnostic kit. The buffer used for enzyme chemistry and HPLC separation of basic drugs is relatively stable, with pKa value of 10.4 and pH buffer range of 9.7-11.1 at 25℃. It is widely used in biochemical analysis and in vitro diagnosis.   CAPS in carton   In addition to the biochemical industry, CAPS is widely used in the following fields:   1. CAPS is widely used as biological buffer: in biochemical diagnostic kit, DNA/RNA extraction kit and PCR diagnostic kit; in enzyme chemistry and HPLC buffer for separation of basic drugs. When caps is used as biological buffer, it needs better purity. Generally, it needs analytical purity. When it is used in industry, the purity requirements are relatively low. However, different treatments should be made for different applications.   2. In industry, CAPS is used for new coatings and materials. CAPS is also the raw material for manufacturing welding materials, air conditioning equipment and lithium metal.   3. It is used as analytical reagent, heat exchange carrier and pharmaceutical industry. It is used for air conditioning, fireworks, dry batteries and lithium metal, as flux and desiccant.   4. For coating industry, it is used as curing agent of water-based isocyanate. The water-based isocyanate curing agent can coexist with resins containing active groups (hydroxyl, carboxyl, amino, epoxy, etc.) for a long time at room temperature.   CAPS has such a wide range of uses and many manufacturers. When selecting manufacturers, attention should be paid to identify qualified manufacturers and avoid intermediaries to make profits from them. Hubei New Desheng Materials Technology Co., Ltd. is located in Guanggu United Science and Technology City C8-2, Gedian Development Zone, Ezhou City, Hubei Province. It has 14 years of research and development and production experience in the field of biochemistry and specializes in the production of biological buffers.The enterprise is ISO 9001 quality management system certification approved and has a good reputation in the industry. It is a trusted manufacturer of biochemical raw materials. It is absolutely wise for you to choose Desheng.

Polyurethane coating is a kind of coating technology which began to rise in the 1960s. In view of the increasingly strict requirements of environmental protection laws and regulations, the environmentally friendly water dispersive polyisocyanate has shown its importance in various application fields in recent years. Although polyether modified polyisocyanates are widely used, they all have one basic disadvantage, especially when they are used as crosslinking agent of waterborne two-component polyurethane coating, higher polyether content is needed to ensure sufficient dispersion, which leads to a long drying time and long-lasting hydrophilicity of the coating. These shortcomings have been solved on the CAPS (3-(cyclohexylamine)- 1-propanesulfonic acid) modified polyisocyanate.           CAPS       CAPS (an amphoteric sulfamate) reacts with aliphatic polyisocyanate under mild conditions and in the presence of tertiary amine neutralizer. The sulfonylurea derivative is an excellent emulsifier. Without considering the influence of salt forming group, CAPS modified polyisocyanate has very good storage stability and the finished product is not turbid. Even if it contains less sulfonate groups, it can also be in water The emulsion with good dispersing state was obtained. Thus, a series of ionic modified polyisocyanates can be obtained, which can be used in various environment-friendly high-quality two-component polyurethane coatings.   These coatings are completely superior to general solvent based coatings in terms of dryness, curing and chemical resistance. With the requirements of new regulations, VOC (volatile organic compounds) content in decoration materials is further reduced, which makes the amount of these cross-linking agents increase greatly. Compared with solvent based coatings, they will not lead to the degradation of film quality. CAPS modified polyisocyanate has been widely used in automobile original paint, repair paint, plastic paint, wood paint, fabric leather paint, printing ink industry, etc. with its excellent performance. The market prospect is self-evident.           Preparation of CAPS modified polyisocyanate       950g polyisocyanate was stirred with 50g CAPS, 29g dimethylcyclohexylamine and 257g 1-methoxypropyl-2-yl acetate under dry nitrogen for 5 hours at 80 ℃, wherein the polyisocyanate is based on hexamethylene diisocyanate (HDI) and contains isocyanurate group, NCO content is 21.7%, average NCO function is 3.5, monomer HDI content is 0.1% and viscosity is 3000mpas. After cooling to room temperature, colorless and transparent polyisocyanate solution can be obtained.           CAPS produced by Desheng company has not only been widely used in the new paint market, but also in the field of biochemical industry. Because it is also a biological buffer, it is widely used in biochemical diagnostic kits, DNA/RNA extraction kits and PCR diagnostic kits. Welcome enterprises and individuals to call for turther consultation.      

Introduction   CAPS, 3-(cyclohexylamine)-1-propanesulfonic acid, an organic chemical, white crystalline powder, CAS1135-40-6, is a biological buffer, commonly used in biochemical diagnostic kits, DNA/RNA extraction kits and PCR diagnostic kits. Buffer for enzyme chemistry and HPLC separation of basic drugs is widely used in the membrane transfer process of protei sequencing. CAPS buffer is composed of CAPS, deionized water, etc., and its pH is adjusted to 11.0 for purificationof fibronectin.   CAPS buffer   Key points of operation   1. SDS-PAGE electrophoresis: according to the conventional conditions (CAPS system: for > = 20kD protein; Tris Tricine system: for low molecular weight protein, also for high molecular weight protein); 2. Methanol concentration: the range of methanol concentration in CAPS electroimprinting buffer is 0-20% (methanol concentration is high, used for low molecular weight protein transfer; methanol concentration is low or even does not contain methanol, used for high molecular weight protein transfer); 3.PVDF membrane treatment: take out the PVDF membrane, soak it in methanol for several seconds, and then put it into the CAPS electroblotting buffer. (Note: the PVDF membrane shall be prevented from drying up after operation. If the membrane is dry, repeat the operation of this step); 4. Gel treatment: remove gel gel and soak in CAPS buffer for 5-10 minutes. (Note: this step can be omitted when transferring some strong basic proteins PI > 9.0); 5. Install the transfer slot: immerse the filter paper and sponge in the electro blotting buffer, then press the electrical imprinting sandwich in the order of sponge, filter paper, PVDF film, gel, filter paper and sponge, and put it into a small electric rotary slot. 6. Transfer conditions: transfer at 50V constant pressure (100-170 MA) at room temperature for 0.5-2h. (Note: drain the bubbles between the gel and PVDF film. For example, the protein above 70 KD should be transferred for longer time); 7. Pretreatment of PVDF membrane dyeing: take out PVDF membrane and rinse it with deionized water, soak it in methanol for several seconds, then dye it; 8. Membrane dyeing: Coomassie brilliant blue dyeing (dissolve 0.1% Coomassie brilliant blue R-250 in 40% methanol/1% acetic acid) for 30-50 seconds (not more than 1 minute), decolorizing with 50% methanol (change decolorizing solution frequently), washing with deionized water fully, and then drying;   CAPS buffer preparation   1. 10×CAPS(100mmol/L) buffer solution is prepared as follows: CAPS, 22.13g; add deionized water to 900ml; adjust the pH value to 11.0 with 2mol/L NaOH (about 20ml), then dilute to 1L, and store at 4℃. 2. CAPS electroimprinting buffer (1×CAPS containing 10% methanol) is prepared by the following methods: 200ml of 10 × CAPS; 200ml of methanol; 1600ml of deionized water.   Other applications   CAPS is also used to manufacture welding materials, air conditioning equipment and raw materials for manufacturing; it is also used to manufacture pyrotechnics; it is used as analytical reagent, heat exchange carrier, and also used in pharmaceutical industry; it is used for air conditioning, pyrotechnics, dry batteries; it is also used as flux and desiccant; it is used for curing agent of aqueous isocyanate in paint industry. Desheng company specializes in R&D and production of various biological buffers, including CAPS, Tris, Bicine, MOPS, etc., with high purity ≥ 99%, stable process, welcome enterprises and individuals to further consultation.

Trimethylolminomethane-Tris base is a kind of buffer which is widely used in the field of biochemistry. Its CAS number is 77-86-1. It has the advantages of stable property, not participating in biochemical process and strong buffering ability. It is widely used in the detection of protein and nucleic acid.   Due to the wide application of Tris, some details need to be noted. Although Tris buffer has strong buffering capacity, it needs to be used carefully when the dilution is too large or the volume of reaction system changes greatly. When the dilution is ten times, the pH change is greater than 0.1, and the pH change of phosphate buffer is less than 0.1.   When preparing nucleic acid agarose gel electrophoresis, the first preparation work is to prepare gel. The quality of agarose gel directly influences electrophoresis efficiency and efficiency. This process takes a long time and saves time to purchase the prepared gel directly.   After the electrophoretic solution is prepared, it can be put into the electrophoresis tank, start sampling, and then turn on the power supply to start electrophoresis. Using TAE/TBE, it generally takes 20-30 minutes to complete electrophoresis, and the voltage is recommended not to exceed 200V. The voltage of TAE is relatively higher than that of tbe electrophoretic solution. The higher the voltage, the faster the electrophoresis rate, but the corresponding resolution will be lower.   The conductivity of tbe is higher than that of TAE. Therefore, compared with TAE, tbe is less likely to cause overheating in the electrophoresis tank, so tbe is recommended for long-term electrophoresis. Boric acid is an inhibitor of enzyme, so if you need to separate electrophoresis DNA for enzyme cutting reaction, TAE is recommended as electrophoresis buffer solution. TAE is better for the separation of longer fragments, and tbe is suitable for fragments less than 300 BP. TAE is more suitable for the recovery of DNA fragments.   Tris, like bicine, is a biological buffer developed and produced by Desheng. In addition, it also has rich production experience in EDTA, virus transportation medium and nucleic acid extraction solution associated with it. Looking forwards to your further consulation.

Tris-trihydroxymethylaminomethane is an organic compound with the molecular formula of (hoch2) 3cnh2. Tris is commonly used in the Tae and tbe buffers (for the dissolution of nucleic acids) in biochemical experiments. Its amino group can condense with aldehydes.   Tris is a weak base, and the pH value of Tris alkali aqueous solution is about 10.5. Generally, hydrochloric acid is added to adjust the pH value to the required value to obtain the buffer solution of this pH value. The effective buffer range of the buffer solution is between pH7.0 and 9.2, but the effect of temperature on the pKa of Tris should be noted. At room temperature 25 ℃, PKA is 8.1.   Because Tris buffer is a weak alkaline solution, DNA will be deprotonated in such a solution to improve its solubility. People often add EDTA into Tris hydrochloric acid buffer to make "Te buffer", which is used to stabilize and store DNA. If the acid solution regulating pH value is replaced with acetic acid, then "TAS / acetate / EDTA" is obtained, and "Tris / borate / EDTA" is obtained when the acid solution is replaced with boric acid. These two buffers are usually used in nucleic acid electrophoresis experiments.   Preparation of Tris HCl and Tris EDTA: 1、 1m Tris HCl (pH 7.4, 7.6, 8.0) to prepare 1000ml Preparation method: a. Weigh 121.1g Tris into a 1000ml beaker. b. Add about 800ml deionized water, and fully stir to dissolve. c. Add concentrated HCl into the following table to adjust the required pH value. pH value Concentrated HCl 7.4 About 70ml 7.6 About 60ml 8.0 About 42ml   d. Dilute the solution to 1000ml. e. Store at room temperature after high temperature and high pressure sterilization.   Note: the solution should be cooled to room temperature before setting the pH value, because the pH value of Tris solution varies greatly with the temperature. The pH value of the solution will be reduced by about 0.03 units for every 1 ℃ rise in temperature.   2、 1.5m Tris HCl (pH 8.8) to prepare 1000ml Preparation method: a. Weigh 181.7g Tris into a 1000ml beaker. b. Add about 800ml deionized water, and fully stir to dissolve. c. Adjust the pH value to 8.8 with concentrated HCl. d. Dilute the solution to 1000ml. e. Store at room temperature after high temperature and high pressure sterilization.   Note: the solution should be cooled to room temperature before setting the pH value, because the pH value of Tris solution varies greatly with the temperature. The pH value of the solution will be reduced by about 0.03 units for every 1 ℃ rise in temperature.   3、 TE is Tris EDTA buffer (10mm Tris, 1mm EDTA, pH7.4, ph7.6, ph8.0). 10 × TE buffer (pH 7.4, 7.6, 8.0) 100mm Tris HCl, 10mm EDTA to prepare 1000ml Preparation method: a. Measure the following solutions and put them into a 1000ml beaker. ①1M Tris-HCl Buffer(pH7.4，7.6，8.0) 100ml； ②500mM EDTA(pH8.0) 20ml b. Add about 800ml deionized water into the beaker and mix evenly. c. After the solution is fixed to 1000ml, it is sterilized under high temperature and pressure. d. Store at room temperature.   Due to the wide application of Tris buffer, in order to highlight the advantages of Desheng company in biological buffer products, we strictly check all aspects of R&D, production and sales, and strive to give the best products to consumers, so that everyone can use them safely, easily and effectively.

  Operation process of Hanks virus transportation medium: （1） Accurate weighing of reagents: select appropriate culture medium according to different fungi and uses, and the reagents required for the culture medium must be pure.   （2） Correction of pH value: put various components of the weighed culture medium into the container, mark and draw lines, heat and dissolve, supplement water, and determine the pH value. It is usually measured by a precision test paper or pH meter with pH of 6.8-8.0. Use 1N NaOH and 1N HCl to adjust the pH value to a suitable range.   （3） Filtration: put the glass funnel on the iron frame, then use gauze with cotton or filter paper to put it in the funnel, pour the above medium into it and filter until it is transparent.   （4） Subpackage: subpackage the filtered culture medium into the medium test tube or triangle bottle (5ml for each tube; 100ml to 150ml for each triangle bottle), pack the cotton plug and wrap it with kraft paper for sterilization.   （5） Sterilization: medium sterilization, commonly used high-pressure steam sterilization. Generally, the nutritional cells of microorganisms are killed when they are boiled in water, but the spores of bacteria have strong heat resistance, so they must be sterilized by high pressure steam to achieve the goal of thorough sterilization. According to the principle that the steam temperature increases with the pressure, that is, the higher the pressure, the higher the steam temperature. Therefore, under the same temperature, high pressure steam sterilization is better than dry heat sterilization. Moreover, in the case of damp heat, the protein is easy to coagulate and denature after the bacteria absorbs water, because the steam has strong penetration and good sterilization effect.   Hanks virus transportation medium     Attention 1. Samples of Hanks virus transportation medium should be detected when they are fresh. In case of bacterial contamination, the bacteria may contain endogenous HRP, and may also produce false positive reaction. If stored for a long time, it can polymerize in ELISA to deepen the background. 2. After the frozen sample is dissolved, the protein is partially concentrated and unevenly distributed. It should be fully mixed, gently and slowly, to avoid bubbles. It can be mixed upside down, and it should not be strongly shaken on the mixer.   3. Turbid or precipitated samples should be centrifuged or filtered first, and then tested after clarification.   Repeated freezing and thawing will reduce the titer of the protein, so if the sample to be tested needs to be preserved for multiple tests, it should be stored in a small amount of sub packed ice. There are some reasons for the straightness of the standard curve in ELISA: whether the preparation of the standard curve is improper, whether the hole washing part is sufficient, whether the reading is inaccurate or whether there is suction error. Find the reason and find the solution.   Storage conditions Due to the different raw materials and requirements, the storage of the medium is slightly different. The general culture medium is easy to be polluted or decomposed by bacteria after being heated and hygroscopic. Therefore, the general culture medium must be kept in a damp proof, dark and cool place. For some culture media (such as tissue culture media) that need strict sterilization, they must be stored in a refrigerator of 2~6℃ for a long time. Because the liquid culture medium is not easy to keep for a long time, it has been transformed into powder.   The Hanks virus transportation medium independently R&D by Desheng has been successfully put on the market, and customers in need are welcome to inquire for details.

Buffer, as a type of substance that can maintain the pH stability of the reaction system, is usually composed of weak acids, weak bases, and their salts or zwitterionic substances. In biological systems, they play crucial roles, such as CO ₂ in photosynthesis and bicarbonate in human plasma. In the field of biochemical testing, Tris buffer and phosphate buffer (PBS) are the two most commonly used, although they each have their own characteristics and application ranges.   Firstly, from the perspective of buffering range, Tris buffer typically has a pH range of 5.0 to 9.2, which varies depending on the acidity of Tris. In biochemical research, Tris buffer has been paid more and more attention because of its wide application range, especially in SDS polyacrylamide gel electrophoresis and other experiments, its use frequency even exceeds that of phosphate buffer. Phosphate PBS buffer has a wider buffering range, covering the pH range of 1 to 12. This is due to the dissociation characteristics of phosphate, which makes the prepared buffer have a wider pH adaptability.   Secondly, from the perspective of application fields, Tris, as an amino buffering agent, has a sodium free characteristic that prevents it from introducing sodium and potassium ions into the system, thereby avoiding the impact on osmotic pressure. In addition, Tris has a relatively small impact on biochemical processes and does not form precipitates with calcium, enzymes, and heavy metal ions, making it suitable for DNA, RNA, and protein related experiments. However, the pH value of Tris is temperature sensitive and the solution is prone to absorbing CO ₂, so special attention should be paid when using it.   Although phosphate buffer has a slightly weaker buffering ability under alkaline conditions, it can dissociate cations and has a salt balance effect, with little impact on the protein structure and biological characteristics of living organisms. Therefore, phosphate PBS buffer is often used in cell experiments. However, it may also form precipitates with calcium, magnesium ions, and heavy metal ions, thereby inhibiting the activity of certain enzymes.   Overall, Tris buffer and phosphate buffer each have their unique advantages and applicability. With the deepening of biochemical research, the application of Tris buffer is becoming increasingly widespread, and there is a trend of gradually surpassing phosphate buffer. However, when choosing which buffer to use, it is still necessary to consider the specific experimental requirements and conditions comprehensively.Desheng specializes in the production of biological buffering agents, with a large quantity in stock. Welcome to consult and purchase!

Trimethylolaminomethane Tris, namely aminobutriol, also known as bradycardic acid amine, is a kind of ternary alcohol containing amino group, which has weak alkalinity. It is usually used with weak acid as Good’s buffer and widely used in biochemical detection and kits in biochemistry and molecular biology.   Good's buffer Tris powder   Physical and chemical properties of Tris English name: Trometamol Molecular weight: 121.135 Molecular formula: (HOCH2)3CNH2 Appearance: white crystal powder Solubility: soluble in water and ethanol, slightly soluble in ethyl acetate and benzene, insoluble in ether and carbon tetrachloride. pKa: 8.1 at room temperature, pH10.5 in aqueous solution Buffer range: 7.0~9.2 Tris buffer is usually used as a solvent for nucleic acids and proteins. Because the carbon atom at position 2 of Tris is connected with amino group, and the aqueous solution is alkaline, when DNA dissolves in this solution, it will be deprotonated, so as to improve the solubility. As a buffer, Tris is usually used with weak acid, such as hydrochloric acid or Tris HCl salt. In addition to HCl, it is often used with acetic acid and boric acid, as well as glycine in electrophoresis buffer.   Tris HCl buffer Weigh the required mass according to the molecular weight of Tris (commonly used concentration is 1~2M), prepare an aqueous solution, and then slowly add concentrated hydrochloric acid to adjust to the required pH value. Note that when the prepared solution is alkaline, it will absorb the acid gas CO2 in the air, and the bottle cap needs to be closed tightly; when adjusting the pH, the solution needs to be cooled to room temperature, and the solution is sensitive to temperature. When the pH value is increased by 1℃, the pH value will drop by 0.03, otherwise, it will change when it is cooled to room temperature after adjusting the pH value.   TE buffer Tris EDTA buffer, commonly used in molecular biological reagents, dissolves DNA. The commonly used concentration is 10-100mM, and the molar concentration of EDTA is one tenth of it. Hydrochloric acid regulates the pH to the required value.   TAE buffer: TrisAcetateEDTA, where acetic acid is used instead of hydrochloric acid.   TBE buffer: Tris+Borate+EDTA, adjust pH and replace hydrochloric acid with boric acid.   Tris-Gly buffer: adjust pH and replace hydrochloric acid with glycine. TBS buffer: In addition to Tris base, salt NaCl and a small amount of KCl should be added to the solution, and the pH should be adjusted with concentrated hydrochloric acid   TBST buffer: add 0.1% Tween 20 to TBS.   Besides being used as DNA, protein lysis buffer, electrophoretic buffer and SDS-PAGE gel electrophoresis, Tris can also react with carbonic acid as humic acid amine in body fluid, generate bicarbonate, absorb hydrogen ions and correct acidemia, and have strong action and can penetrate cell membrane. Tris produced by Desheng is mainly used for Good’s buffer and mass export for further refining.

PEP, namely phosphoenolpyruvate, is a very important substance in all life activities. It participates in many biochemical processes such as cell respiration and photosynthesis. The reagent we produce refers to its salt, phosphoenolpyruvate tricyclohexamine salt reagent.   Physical and chemical properties of PEP salt English name: Phosphoenolpyruvic acid tricyclohexylammonium salt Molecular weight: 465.56 Molecular formula: C21H44N306P Molecular structure   Application of serum carbon dioxide determination kit In the chloroplast, PEP can absorb carbon dioxide and convert it to oxaloacetate under the catalytic action of PEP carboxylase (carboxykinase). This process is the core of photosynthesis. The efficiency of PEP carboxylase to fix CO2 is very high, and the reaction is very sensitive. With this feature, the content of trace carbon dioxide in serum can be determined, and the activity of PEP carboxylase in plant samples or serum or plasma can also be measured.                                                                CO2 fixation process of PEP in photosynthesis   Determination principle PEP and bicarbonate (the form of serum CO2) are catalyzed by enzyme to produce oxaloacetate. Oxaloacetate and NADH are catalyzed by malate dehydrogenase MDH to produce malate malate. NADH (nicotinamide) is measured with a spectrophotometer The reduced state of adenine dinucleotide, reduced coenzyme I) The absorbance at 340nm is attenuated, and the amount of attenuation is proportional to the concentration of CO2, thereby measuring the serum CO2 content. Similarly, the enzyme activity can be measured according to the rate of change of absorbance when a certain amount of CO2 is generated, that is, a reagent kit for measuring the activity of a sample PEP carboxylase.   PEP is used in cell protectants and antioxidants In cellular respiration, PEP participates in the last step of glycolysis, which is converted to pyruvate after removing high-energy phosphate groups. In the process of organ tissue refrigeration, it can reduce the oxidative stress and ATP consumption of tissue cells, reduce organ damage, and prolong the preservation time of clinical organs.   The use of PEP salt is not limited to this. Due to its carbon dioxide absorption and cell glycolysis characteristics, many applications are still under development. Desheng Technology's mature reagent is PEP tricyclohexylamine salt, mainly used Kit for carbon dioxide determination and PEP carboxylase activity determination.

Phosphoenolpyruvate (PEP) is a glycolysis intermediate. Phosphoric acid is a glycolysis metabolite with a high-energy phosphate group. After dephosphorylation, PEP is converted to pyruvate, which can penetrate cell membranes With cell protection and antioxidant activity, it can be used as a cell protection agent and antioxidant in the liver of cold-preserved mice and a potential organ protection agent.   Its cell protection effect is mainly reflected in the following aspects 1. PEP (0.1-10 mM) significantly attenuated the hydrogen peroxide-induced reduction in cell viability in HeLa cells in a dose-dependent manner.   2. PEP also inhibited the decrease of calcein-acetylmethoxy-stained cells and the increase of propidium iodide-stained cells induced by hydrogen peroxide. The increase of intracellular reactive oxygen species stimulated by hydrogen peroxide was significantly reduced.   3. Evaluation by electron paramagnetic resonance method also shows the potential of PEP to scavenge hydroxyl radicals.   4. In addition, PEP has the potential to scavenge 1,1-diphenyl-2-pyridylmethylhydrazonyl radical (a representative artificial free radical), although the potential is small.   5. PEP (10 mM) slightly inhibited the reduction of H2O2-induced cellular ATP content, but did not show any effect at low doses (0.1, 1 mM).   6. PEP (0.1-10 mM) also reduced cell damage, but did not attenuate the decrease in intracellular ATP content induced by the glycolysis inhibitor 2-deoxy-D-glucose.   These results indicate that PEP exerts a cytoprotective effect and has antioxidant potential, although the exact cytoprotective mechanism has not been fully elucidated. However, we believe that PEP is a functional carbohydrate metabolite with cell protection and antioxidant activity, and may be used as a therapeutic agent against diseases involving oxidative stress.   In addition, the PEP produced by Desheng company can also be used to determine the content of CO2 in the biochemical kit. The content of CO2 reflects the metabolic acid-base balance in the body. It can also be used for the auxiliary diagnosis of diseases such as pyloric obstruction, Cushing's syndrome, taking too many alkaline drugs and respiratory acidosis.

HEPES is a kind of hydrogen ion amphoteric buffer with good buffer ability and long time to maintain pH constant. It is widely used in nucleic acid reaction buffer, hybridization buffer and cell culture medium. According to its characteristics, there are some differences in the configuration of buffer in different experiments.   Physical and chemical properties of HEPES 2-[4-(2-Hydroxyethyl)-1-piperazinyl]ethanesulfonic acid CAS No.: 7365-45-9 Molecular formula: C8H18N2O4S Molecular weight: 238.3 Buffer range: 6.8-8.2 Molecular Structure: HEPES mother liquor (buffer stock): directly prepare 0.5-1m HEPES solution and NaOH solution, control the mixing ratio of the two to adjust the pH, and then use distilled water or ultra pure water to fix the volume. If necessary, NaOH can be replaced by KOH. HEPES buffer salt solution: add a small amount of sodium chloride and disodium hydrogen phosphate into HEPES solution, then add NaOH aqueous solution, adjust pH, add distilled water for constant volume, and store at low temperature.   HEPES cell culture medium: add a small amount of sodium chloride, potassium chloride, disodium hydrogen phosphate, dextran, etc. into HEPES aqueous solution, then adjust the pH with NaOH solution, and finally store at a constant volume and low temperature. In cell adhesion experiments, calcium and magnesium ions are usually added to HEPES culture medium to protect the cadherin of cells and not affect the formation of cell aggregation.   HA solution: HEPES-BSA cell culture medium, which is one of the components of cell culture medium, does not contain calcium and magnesium ions, and contains some other salt ions. After the treatment of filtration bacteria, it is mostly suitable for cleaning and culture in primary culture of tissue cells.   The above is the application difference of HEPES buffer developed and produced by Desheng in different experiments. In addition, the non inactivated virus preservation solution newly developed by the company also contains HEPES buffer. Because it has no toxic effect on cells, it not only maintains pH, but also increases the in vitro survival time of virus host cells after sampling, retains the integrity of virus nucleic acid and protein to the greatest extent, and improves the detection accuracy.