Dear PANKAJ BHATT

HEPES buffer which was first described by Good, et al. (1966). acts as a zwitterions, proved superior to conventional buffers in comparative biological assays with cell-free preparations. It has many properties which make it ideal as a buffer to tissue culture media, it does not require an enriched atmosphere to maintain the correct pH. HEPES does not bind divalent cations and is soluble to the extent of 2.25 M at 0°C. Note: since the DpKa/°C of -0.014, the pH reading recorded in a HEPES buffered medium will vary inversely with the temperature of the medium.

Pls. find the attached files

http://www.microbiologyresearch.org/docserver/fulltext/jgv/2/2/JV0020020309.pdf?expires=1474706991&id=id&accname=guest&checksum=B83D6BC1A2CCFFB3BC8EC766B3ED285D

http://www4.mpbio.com/ecom/docs/proddata.nsf/5f64ffd4f38c2fda8525645d00769d68/d129d700747af023852568b200619fff

http://cellgro.com/media/upload/file/techinfosheets/new/Buffering%20Systems.pdf

https://en.wikipedia.org/wiki/HEPES

https://www.researchgate.net/post/Is_it_important_to_add_HEPES_to_primary_cell_cultures_for_proliferation_assays

Hoping this will be helpful            

Regards

Prof. Houda Kawas

Dear PANKAJ BHATT

HEPES buffer which was first described by Good, et al. (1966). acts as a zwitterions, proved superior to conventional buffers in comparative biological assays with cell-free preparations. It has many properties which make it ideal as a buffer to tissue culture media, it does not require an enriched atmosphere to maintain the correct pH. HEPES does not bind divalent cations and is soluble to the extent of 2.25 M at 0°C. Note: since the DpKa/°C of -0.014, the pH reading recorded in a HEPES buffered medium will vary inversely with the temperature of the medium.

Pls. find the attached files

http://www.microbiologyresearch.org/docserver/fulltext/jgv/2/2/JV0020020309.pdf?expires=1474706991&id=id&accname=guest&checksum=B83D6BC1A2CCFFB3BC8EC766B3ED285D

http://www4.mpbio.com/ecom/docs/proddata.nsf/5f64ffd4f38c2fda8525645d00769d68/d129d700747af023852568b200619fff

http://cellgro.com/media/upload/file/techinfosheets/new/Buffering%20Systems.pdf

https://en.wikipedia.org/wiki/HEPES

https://www.researchgate.net/post/Is_it_important_to_add_HEPES_to_primary_cell_cultures_for_proliferation_assays

Hoping this will be helpful            

Regards

Prof. Houda Kawas

Dear Pankaj Bhatt, Perfect answer above. HEPES is a robust buffer in tissue culture, and it does not require CO2 enriched atmoshpeher, however, some mammalian primary cells/ cell lines may not like it due to its albeit low toxicity level. Regards, -Laszlo Takacs

Dear Pankaj,

HEPES : - A biological buffer (exist in a zwitterionic form ). It maintains physiological pH in cell culture when compared to bicarbonate buffers despite the changes in carbon dioxide concentration. i suggest you to go through the below mentioned link for more information. 

http://www4.mpbio.com/ecom/docs/proddata.nsf/5f64ffd4f38c2fda8525645d00769d68/d129d700747af023852568b200619fff

Thanks,

Prakash

HEPES is a zwitterionic organic chemical buffering agent; one of the twenty Good's buffers. HEPES is frequently added to cell culture media in applications when pH maintenance is critical and normal bicarbonate buffering is not sufficient. It has no nutritional benefit to cells. It is added to the media solely for extra buffering capacity when cell culture requires extended periods of manipulation outside of a CO2 incubator.

Thanks to all of you