HEPES Buffer CAS 7365-45-9
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Factory Supply HEPES Buffer CAS 7365-45-9 with Best Price
- Appearance: Colorless liquid
- Purity: 99. 0%min
- Stock: In stock
- Sample: Available
- Zhishang Chemical: HEPES buffer Supplier & Manufacturer
HEPES Buffer: The Complete Guide
- Item 1: Basic Info of HEPES Buffer
- Item 2: What is HEPES Buffer?
- Item 3: HEPES Buffer Uses
- Item 4: The Manager has Something to Say
- Item 5: Application of HEPES Buffer
- Item 6: Preparation of HEPES Buffer
- Item 7: Quality Control of HEPES Buffer
- Item 8: Hot Sale Products
- Item 9: Get HEPES Buffer Quotation
HEPES Buffer on Sale
Basic Info of HEPES Buffer
Basic Info
Chemical Properties
Other Info
Basic Info
Chemical Name: | HEPES Buffer |
Other Name: | HEPES，Free acid; HEPES, Biological Buffer, Ultrapure; 2-(4-(2-Hydroxyethyl)piperazin-1-yl)ethanesulfonic acid; 4-Hydroxyethylpiperazine ethanesulfonic acid |
CAS: | 7365-45-9 |
EINECS: | 230-907-9 |
Type: | Pharmaceutical raw materials; Pharmaceutical intermediates; Dye intermediates; Pesticide intermediates; Cosmetic raw materials |
HEPES Molecular Formula: | C8H18N2O4S |
HEPES Molecular Weight: | 238.3 |
Chemical Properties
Melting point | 234-238 °C |
density | 1.07 g/mL at 20 °C |
refractive index | n20/D 1.339 |
storage temp. | 2-8°C |
solubility | H2O: 1 M at 20 °C, clear, colorless |
form | powder |
color | White |
PH | 5.0-6.5 (1Maqueous solution) |
PH Range | 7.0 – 7.6 |
HEPES pka | 7.5(at 25℃) |
Water Solubility | Soluble |
λmax | λ: 260 nm Amax: 0.06 |
Merck | 14,4654 |
BRN | 883043 |
HEPES Stability: | Stable. Combustible. Incompatible with strong oxidizing agents. Protect from moisture. |
Other Info
Brand Name: | Zhishang Chemical |
Provide: | HEPES Buffer MSDS; HEPES Buffer COA |
What is HEPES Buffer?
HEPES buffer(4 – (2-hydroxyethyl) – 1-piperazine ethanesulfonic acid) is a zwitterionic sulfonic acid buffer; One of good’s 20 buffers. Hepes buffer is widely used in cell culture, mainly because it can maintain physiological pH value better than bicarbonate buffer, which is also commonly used in cell culture, although the concentration of carbon dioxide (produced by aerobic respiration) changes. Lepe Zuniga et al. An unwanted photochemical process in which hepes buffer produces hydrogen peroxide when exposed to ambient light is reported, which is not a problem in bicarbonate based cell culture buffer. Therefore, it is strongly recommended to place the solution containing hepes buffer in the dark as far as possible to prevent oxidation.
HEPES buffer has the following characteristics：
p K a1 (25 °C) = 3
p K a2 (25 °C) = 7.5
Useful pH range = 2.5 to 3.5 or 6.8 to 8.2
The negligible metal ion binding of hepes buffer makes it an ideal buffer for enzymes that may be inhibited by metal chelation.
HEPES Buffer Uses
- HEPES buffer(4-hydroxyethylpiperazineethanesulfonic acid) is a non-ionic amphoteric buffer, highly polar, inert to a variety of chemical reagents and enzymes, does not participate in and does not interfere with the process of biochemical reactions, and is resistant to enzymatic chemical reactions. There is no inhibitory effect on the reaction, so it can be specially used for the research of organelles and highly volatile, pH-sensitive proteins and enzymes.
- Cell-cell adhesion, short-term cell aggregation culture, buffer for washing tissues and cells.
- Patent CN201310477071.6 discloses a method for preparing porous zinc oxide microspheres by using HEPES buffer(4-hydroxyethylpiperazine ethanesulfonic acid) as a template. The method is simple to operate, and the prepared microspheres have the advantages of uniform size and hierarchical pore structure, etc., and can be used for the production of catalysts and gas sensing elements.
- Patent CN201080052103.2 uses HEPES buffer(4-hydroxyethylpiperazine ethanesulfonic acid) and its derivatives to treat cancer pain and suppress the side effects after chemotherapy.
- Hydroxyethylpiperazineethanesulfonic acid is a common component of buffers in animal and human cell cultures. Studies have confirmed that HEPES buffer(4-hydroxyethylpiperazine ethanesulfonic acid) has minimal cytotoxicity among all known buffers. The FDA (US Food and Drug Administration) has approved piperazine-based zwitterionic molecules as food additives.
The Manager has Something to Say
This is William, CEO of Zhishang Chemical Co., Ltd.
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Application of HEPES Buffer
The enhancement effect of 4-hydroxyethylpiperazineethanesulfonic acid(HEPES buffer) on tuna peptide xanthine oxidase inhibitor, the specific experimental steps are as follows:
- Prepare 150mM 4-hydroxyethylpiperazineethanesulfonic acid(HEPES buffer), Tris-HCl, and PB buffer at 37°C with pH 7.4, and store at 4°C until use;
- Prepare the required sample (tuna peptide) with the above-mentioned 150 mM buffer, and the sample concentration is 40 mg/ml;
- Add 50 μL tuna peptide or 50 μL buffer (blank) and 150 μL xanthine in sequence to the 96-well microtiter plate, and do 3 parallels for each sample, incubate at 37°C for 5 min, add 50 μL xanthine oxidase, and read once in 30 s Absorbance value, a total of 50 times 25min. After reading, the reaction was terminated with 80 μL of HCl, and the reaction solution was diluted 10 times with the corresponding buffer and passed through a 0.25 μm aqueous membrane to measure the uric acid concentration;
- Measure the content of uric acid in the sample after the reaction by high performance liquid chromatography, and quantify it with a uric acid standard;
Calculate the inhibition rate of xanthine oxidase, and the calculation method is as follows:
I=(CB-CS)/CB×100%
In the formula: I represents the inhibition rate of xanthine oxidase, CB is the peak area of the uric acid peak in the liquid phase spectrum after the reaction in the blank group (without adding the peptide), and CS represents the peak area of the uric acid peak in the liquid phase spectrum after the reaction in the sample group (with the peptide added).
In this method, the preparation process of tuna peptides is as follows: the tuna fish meat is crushed and mixed with deionized water according to the mass ratio of material to liquid of 1:1, the pH value is adjusted to 7.0, and compound protease and papain are respectively added according to 0.6-1.2% of the mass of tuna. , enzymolysis at 50-60°C for 4-7 hours, after the enzymolysis is completed, the temperature is raised to 95°C for 15 minutes to inactivate the enzyme, cooled, centrifuged, and the supernatant is concentrated and spray-dried to obtain tuna peptides.
In 4-hydroxyethylpiperazine ethanesulfonic acid buffer(HEPES buffer), Tris buffer and PB buffer, the content of uric acid produced by xanthine oxidase catalyzed by xanthine has almost no difference, while HEPES can significantly increase the xanthine content of tuna polypeptide oxidase inhibition rate, this result indicates that 4-hydroxyethylpiperazine ethanesulfonic acid(HEPES buffer) can enhance the inhibitory activity of polypeptide xanthine oxidase.
Preparation of HEPES Buffer
Method 1
Method 2
Method 1
Straight make use of 1,2-dichloroethane as solvent, include hydroxyethylpiperazine (5.00 g, 0.02 mol), potassium carbonate K2CO3 (6.00 g, 0.04 mol) in a 100mL there-necked flask equipped with mechanical mixing and thermostat, 50mL of 1,2-dichloroethane, warmed in an oil bath at 90 ° C( boiling point of 1,2-dichloroethane is 85 ° C), and also mixed for 20h. The reaction was quit, filtered, and the filteringed system salt was cleaned with 200 mL of ethyl acetate (EA). The filtrate was spin-dried to obtain 2.6 g of HEPES buffer strong.
Method 2
Add 11.0 g (84.5 mmol) anhydrous salt sulfite, 27.0 mL (343.6 mmol) dichloroethane, 120mL water, 110mL ethanol, 50mg copper powder in turn to the three-necked flask furnished with magnetic stirrer and also reflux condenser, stir, oil The bathroom was heated up to reflux. After refluxing for 22 h, the response solution was vaporized under decreased stress to eliminate water till all white solids were precipitated. This solid is composed mostly of product, unreacted starting material, as well as salt created. The obtained strong as well as 500 mL of ethanol were added to a 1 L flask, as well as heated to reflux for 40 minutes. Suction filtering while hot, as well as the filtrate was cooled down as well as positioned at 0 ° C overnight. Suction purification and also vacuum drying to get flaky crystals, the yield is 11.40 g, as well as the yield is 81.0%.
Add salt chloroethanesulfonate (15.10 g, 0.08 mol), hydroxyethylpiperazine (9.88 g, 0.075 mol), 60mL water, oil bath to a four-necked flask equipped with a magnetic stirrer, a reflux condenser and a thermostat. Mix the reaction at 105 ° C. As the reaction progresses, the pH of the response solution will go down. 5mol/L NaOH liquid remedy is included dropwise to regulate the pH to be around 9. A total amount of 15mL is added, and the reaction is continued for 5h.
After the reaction was completed, the reaction service was thinned down to 500 mL with water, as well as was related to an ion exchange resin column (concerning 500 g) for desalting filtration. After the reaction option was all put on the column, washed with pure water until the pH of the effluent was 6, and afterwards rinsed with 1 mol/L ammonia water., accumulate the effluent (pH concerning 5 ~ 9) with product areas found by TLC. It was concentrated to 150mL by rotary dissipation, decolorized by including activated carbon, heated and also mixed in an oil bathroom at 110 ° C for 0.5 h, filteringed system, and the filtrate was spin-dried, included with 50mL of ethanol, heated to reflux for 0.5 h, and warm filteringed system. The gotten white solid was 8.63 g after drying out. Glacial acetic acid was added dropwise to the filtrate, the pH was gotten used to 5, cooled at 0 ° C. overnight, filtered, as well as the acquired strong was 2.80 g after drying out. It is 11.43 g in complete with the aforementioned HEPES strong, as well as the return is 64.5%.
The preparation approach of 10mmol/L hepes barrier is as adheres to: Properly weigh 2.383 g of HEPES, include fresh three-distilled water and also dilute to 1L. Filter disinfect and also save at 4 ° C after aliquoting. If it is made use of to add to the cell society tool as a buffer, it is advised to store the medium in the dark.
References
- Baicu SC, Taylor MJ (2002). “Acid-base buffering in organ preservation solutions as a function of temperature: new parameters for comparing buffer capacity and efficiency”. Cryobiology. 45 (1): 33–48.
- Lepe-Zuniga JL, Zigler JS, Gery I (October 1987). “Toxicity of light-exposed Hepes media”. Journal of Immunological Methods. 103 (1): 145.
- Zigler JS, Lepe-Zuniga JL, Vistica B, Gery I (May 1985). “Analysis of the cytotoxic effects of light-exposed HEPES-containing culture medium”. In Vitro Cellular & Developmental Biology. 21 (5): 282–7.
HEPES Buffer Supplier and Manufacturer
As a HEPES buffer supplier and manufacturer with a strict standard product quality system certificate, Zhishang Chemical has long been providing the best HEPES buffer raw materials to customers all over the world.
Over the years, relying on professional team experience and customer-centric team concept to meet customer needs in a timely manner, our company enjoys a high reputation in the world. At present, our company has served more than 6,000 customers and has established long-term cooperative relations with customers from many countries.
If you have a demand for HEPES buffer and related products, please contact our service staff Zhishang Chemical – White directly, and we will provide you with high-quality products at the best price.
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FAQ
Do you accept sample order?
We will make samples before mass production, and after the sample is approved, we’ll begin mass production. Doing 100% inspection during production, then doing random inspection before packing.
What’s your MOQ?
Our MOQ is 1kg. But usually we accept less quantities, such as 100g on the condition that the sample charge is 100% paid.
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The different quantity has different discount.
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You can get free samples for some products, you only need to pay the shipping cost or arrange a courier to us and take the samples. You can send us your product specifications and requests, we will manufacture the products according to your requests.
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1. ≤50kg, Express delivery recommended, usually called DDU service;
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Quality Control of HEPES Buffer
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