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Cell line identity (HEK293-E)

HEK293 cell identity

Best Answer Osibisa, 18 January 2014 - 06:36 PM

Well as a partial answer to my own question, the STR profile of HEK-293 C18 is available, which is the ATCC's version of HEK-293E. There is a good database at dmsz.de which shows that there are basicaly two very similar profiles for all HEK-293 cell derivatives, so they can't be distinguished.

 

I would need to either western blot with an EBNA-1 antibody or PCR the sequence out, however a positive control would be desirable.

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#1 Osibisa

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Posted 14 January 2014 - 07:31 PM

Hey everyone,

 

After working on AD-HEK293 and HEK293-T cells I switched to HEK293-E. Since these are no longer commerically available I received a sample from another lab. The only problem is my transfection yields haven't increased, although I expected improvement due to use of a new vector with an Eppstein Barr origin of replication. I have suspisions that they might not be true HEK-E cells... but how do I check?

 

Clearly STR profilling is the standard method, which is offered at the labs we get our sequencing performed, but will it discriminate between variants of HEK293? It appears that the profile of HEK293 and HEK293-T differ slightly but I can't find a profile for HEK-E. Does anyone know why they are no longer available for purchase? Were they originally ATCC, then Invitrogen (Life tech)? I take it they have the profile on record.


Edited by Osibisa, 14 January 2014 - 07:31 PM.


#2 Osibisa

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Posted 18 January 2014 - 06:36 PM   Best Answer

Well as a partial answer to my own question, the STR profile of HEK-293 C18 is available, which is the ATCC's version of HEK-293E. There is a good database at dmsz.de which shows that there are basicaly two very similar profiles for all HEK-293 cell derivatives, so they can't be distinguished.

 

I would need to either western blot with an EBNA-1 antibody or PCR the sequence out, however a positive control would be desirable.







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