Hepes(Cell Culture Grade),IC-1329
Product Introduction
HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) is a zwitterionic organic chemical buffering agent widely used in cell culture, molecular biology, and biochemical research. As one of Good''s buffers, HEPES maintains a stable pH range of 6.8 to 8.2 with a pKa of 7.3 at 25¡ãC, providing superior buffering capacity at physiological pH compared to bicarbonate-based buffers. This product is supplied as a ready-to-use 1M sterile solution at pH 7.2-7.4 or as a high-purity dry powder, both manufactured under animal-free conditions with low endotoxin levels specifically validated for cell culture applications. HEPES is particularly valuable for maintaining pH stability in cell culture media during prolonged incubations outside CO₂ incubators, during cell handling procedures, and for experiments requiring precise pH control independent of CO₂/bicarbonate equilibrium. It is also commonly used in protein purification, enzyme assays, and various molecular biology protocols.
Product Features
1. Superior Physiological Buffering: Maintains stable pH between 6.8 and 8.2 with pKa of 7.3 at 25¡ãC, ideal for mammalian cell culture at physiological pH.
2. Animal-Free and Low Endotoxin: Manufactured under animal component-free conditions with certified low endotoxin levels, suitable for sensitive primary cell culture and GMP-compliant workflows.
3. High Purity Grade: ¡Ý99.5% purity (dry powder) ensures minimal interference with cellular processes and biochemical assays.
4. Ready-to-Use 1M Solution: Sterile-filtered liquid formulation at pH 7.2-7.4 eliminates preparation time and pH adjustment steps.
5. Versatile Applications: Compatible with a wide range of cell culture media formulations, protein and enzyme buffers, and molecular biology protocols.
Specifications
Size: Available as 100 mL 1M Solution (pH 7.2-7.4), 25 g Dry Powder, and 100 g Dry Powder
CAS Number: 7365-45-9
Molecular Weight: 238.30
Molecular Formula: C8H18N2O4S
Purity: ¡Ý99.5% (dry powder)
Grade: Animal-Free, Low Endotoxin, Cell Culture Tested
Solubility: Water: 1M (238.3 mg/mL)
Storage and Stability
Storage Conditions: Store at room temperature (15-25¡ãC) or 4¡ãC. Protect from light for long-term storage. The 1M solution is sterile-filtered and should be handled aseptically after opening.
Shelf Life: The product is stable for 36 months from the date of manufacture when stored as directed. After opening, the 1M solution is stable for up to 6 months when stored at 4¡ãC and handled aseptically.
Protocol (For Reference Only)
Important: For cell culture use, add HEPES to culture medium at a final concentration of 10-25 mM. The ready-to-use 1M solution is sterile-filtered; handle aseptically to maintain sterility.
1. Cell Culture Medium Supplementation (1M Solution): Add 1M HEPES Solution directly to complete cell culture medium at a 1:40 to 1:100 dilution to achieve a final concentration of 10-25 mM (e.g., add 0.5-1.25 mL per 50 mL medium). Mix thoroughly by gentle swirling.
2. Preparation from Dry Powder: Dissolve the appropriate amount of HEPES dry powder in ultrapure water or buffer to achieve the desired final concentration, adjust pH using NaOH or HCl as required, filter sterilize (0.22 µm) for cell culture applications, and store at 4¡ãC.
3. Use for pH Maintenance Outside CO₂ Incubator: For prolonged cell handling outside the incubator (e.g., cell sorting, microscopy, or microinjection), supplement the medium with 25 mM HEPES to maintain physiological pH in ambient air conditions.
4. Protein Purification and Enzyme Assays: Prepare HEPES buffers at concentrations of 10-100 mM with appropriate salt and cofactor supplements for protein extraction, chromatography, or enzymatic reaction buffers. Adjust pH to the optimal range for the specific protein or enzyme being studied.
5. Molecular Biology Applications: Use HEPES at 10-50 mM in various molecular biology buffers including hybridization buffers, transfection mixes, and nucleic acid storage solutions where precise pH control is required.
Precautions
1. HEPES is not a primary buffer system in standard bicarbonate-based media; it is used as a supplemental buffer. Do not replace the bicarbonate/CO₂ system with HEPES alone for long-term culture.
2. The 1M HEPES Solution is sterile-filtered; handle aseptically and avoid contamination after opening. Use within 6 months of opening when stored at 4¡ãC.
3. For the dry powder, dissolve completely before pH adjustment. HEPES will not buffer effectively until fully dissolved and pH-adjusted.
4. HEPES can be cytotoxic at concentrations exceeding 50 mM for certain cell types. Titrate the optimal concentration for sensitive primary cells or novel cell lines.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the recommended final concentration of HEPES in cell culture medium?
A1: 10-25 mM HEPES is commonly used as a supplemental buffer in cell culture media. For prolonged cell handling outside the CO₂ incubator, 25 mM is recommended. Do not exceed 50 mM as it may exhibit cytotoxicity in sensitive cell types.
Q2: How is HEPES different from the bicarbonate/CO₂ buffer system?
A2: The bicarbonate/CO₂ system is the primary physiological buffer system requiring a CO₂-enriched atmosphere. HEPES provides additional pH buffering independent of CO₂, making it useful for short-term handling of cells outside the incubator. It is a supplemental, not a replacement, buffer.
Q3: Does this product contain any animal-derived components?
A3: No. This HEPES product is manufactured under animal-free conditions and is tested for low endotoxin levels, making it suitable for sensitive cell culture applications and GMP-compliant workflows.
Q4: Can I autoclave HEPES solutions?
A4: Autoclaving is not recommended for concentrated HEPES solutions as it may cause degradation and discoloration. Use sterile filtration (0.22 µm) to sterilize HEPES solutions prepared from dry powder.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of HEPES concentration is recommended for specific cell types and experimental conditions.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-1329
Product Name: HEPES
Size: 1M Solution, 100 mL / Dry Powder, 25 g / Dry Powder, 100 g
Price:
1M Solution, 100 mL: CNY ¥550.00 / USD $55.00 / EUR €66.00 / JPY ¥9900.00
Dry Powder, 25 g: CNY ¥800.00 / USD $80.00 / EUR €96.00 / JPY ¥14400.00
Dry Powder, 100 g: CNY ¥2600.00 / USD $260.00 / EUR €312.00 / JPY ¥46800.00
2023 Version
