HEPES Stock Buffer Solution

Description

Background

HEPES Stock Buffer Solution is one of the commonly used buffers in cell cultures as well as biochemical analysis. HEPES maintains physiological pH despite changes in carbon dioxide concentration, therefore it acts better than bicarbonate in cell culture applications.  However, HEPES shows an unwanted photochemical process in the presence of riboflavin when exposed to ambient light to produce hydrogen peroxide. We therefore recommend keeping the mentioned solutions in the dark.

Features

• Cell Culture Suited
• Provides a buffer solution to maintain the physiological pH despite changes in carbon dioxide concentration.
• PurMa Biologics manufactures this buffer  using Mg²⁺ and Ca ²⁺free water
• Free of RNAse /DNase
• Free of Proteases. It is rigorously tested for contaminating nonspecific endonuclease, exonuclease, and RNase activity.
• We rigorously test for contamination of endotoxin and mycoplasma.
• PurMa Biologics manufactures this buffer in several PH formats and NaOH adjusts the pH.
• We Sterilize this buffer using  0.2µm filters.
• As it is packed under N_2, the pH won’t change over time before opening the bottle.

PurMa Biologics Manufactures Several Grades of Buffers

Regular Grade Buffer

PurMa Biologics manufactures Regular grade of buffer by using:  PurMa™ Cell Culture Suited Water (Cat# P3W110101)

We sterilize the buffers by autoclaving the solution. Finally we pack the buffer in our Class 100 Aseptic Facility.

Cell Culture Suited Grade Buffer

PurMa Biologics manufactures this grade of buffer by using :PurMa™ Cell Culture Suited Water (Cat# P3W110101)

After adjusting the pH, we finally, sterilize the buffer by  3 times passing through 0.2 µm filters. Additionally,  We pack the buffer under N2 gas  in our Class 100 Aseptic Facility.

RNase and DNase Free Grade Buffer

PurMa Biologics manufactures this grade of buffer by using :PurMa™ Ultra-Pure Cell Culture Suited Water (Cat# P3W110102)

We sterilize the buffers by autoclaving the solution.  Additionally, we pass the buffer  three times through 0.1 µm filters,  and finally  pack it under N2 gas in our Class 100 Aseptic Facility.

Cell Culture Suited / Endotoxin Depleted Grade Buffer

PurMa Biologics manufactures this grade of buffer by using: PurMa™ Ultra-Pure Cell Culture Suited Water (Cat# P3W110102)

After depleting  the endotoxins (see below for more details), we sterilize the buffer by 3 times passing through  0.1 µm filters and pack under N2 gas in our Class 100 Aseptic Facility

The Endotoxin Removing Procedure from Buffer

PurMa Biologics has the most sophisticated endotoxin removal procedure. To remove this pathogen, we pass the buffer  through the PurMa™ Endotoxin Elimination Column (Cat# P6H1216251). Furthermore, in our proprietary column,  we covalently conjugate Sepharose 6B  to an endotoxin-specific absorbent. We finally  eliminate the endotoxin residual by passing the unbound fraction three times through 0.1µm filters.

References

1. Toxicity of light-exposed Hepes media. Lepe-Zuniga JL, Zigler JS Jr, Gery I. J Immunol Methods. 1987 Oct 23;103(1):145. doi: 10.1016/0022-1759(87)90253-5. PMID: 3655381.
2. Analysis of the cytotoxic effects of light-exposed HEPES-containing culture medium. Zigler, JS Jr , Lepe-Zuniga JL, Vistica B, Gery I. In Vitro Cell Dev Biol. 1985 May;21(5):282-7. doi: 10.1007/BF02620943. PMID: 4019356.

 

Learn more: What Is HEPES Buffer?