Hi, Matti Ullah.

Considering all the aspects you're presenting, the first thing I would say is: Congrats!!! You discovered other papers where wrong or the cell line has changed overtime. Check it out!

Have you checked in the ATCC website If they really say this cell line does NOT express the protein you're testing? If so, you may have found something unique.

Other thing for you to check ... Are you using the same protocol (buffers, media, pH, etc) of the papers you looked and that said this protein was negative in this cell line? If not, check it and do as they said they have done and compare it.

One last advice, take care and check If you have not contaminated your cell stocks. Maybe you got a mixed cell stock and now the cells in control and test experiments are quite the same.

Hope it helps.

Hi,

You can perform karotyping to confirm the cell line and also IF for specific gene expression mentioned in ATCC website to confirm the cell line. If tissue specific cell lysate is available commercially, you try that as positive control along with your sample cell lysate and check for the gene expression. Based on the expression from the positive control, you could draw the conclusion.

Thanks

Nisha.P.R

Thanks for your answers.. and they are really helpful.

As far as culture parameters are concerned, yes there is a difference as I used IMDM, but the others used RPMI(dont know why as ATCC recommends IMDM).... Perhaps that might be the issue, although it doesnt make much sense.. ATCC does not specify about this protein so no help from there.

About cell stock contamination, as I have purchased and solo handed the cell line, with no other non-adherent cell line in my experiments, the chances of cross-contamination are nearly zero..

Taking a lysate will be quite a good idea... i will for sure try that.

Thanks