Hi everyone,
does anyone has already tested good working pAkt-Thr308, ALK, pALK-Tyr1604 pBIM-Ser69 TERT and PTERT-Ser824 antibodies for westen blot on human cell lines? If yes, would you be kind as to idicate me from which company did you get them? the ones I am testing now don´t give any good result. I would be glad if you can tell me if one of these have worked in your hands and how. Thanks
Among antibodies you mentioned, I worked with AKT with human cell lines and it works just fine with me. almost 90% of antibodies we are using in our lab we order it from cellsignalling (which is my favorite)
Now, regarding your antibodies, no matter from where you ardered, it doesnt make since to me that none of them is working, There must be somthing in your protocol or simply they are not expressed in your cells, I dont know
Either you should try positive control or run GAPDH or BACTIN in same membrane with any of them to make sure your protocol is fine
I would also prefer if you show any pictures of your results so I can see where is the defect exactly
Thank you For answer Abdullah.
My housekeeping gene works fine. As for AKT, some are working and some not. I am talking about the phosphorylated form and not the total protein. One of my colleague is actually using the phosphoAKT ser473 and he has good results and I am testing phospho-AKT Thr308 and I don´t have good results. beside all the antibodies cited above, I am using oder antibodies, among them Erk, pErk, Bcl2, pbcl2, AKT (total protein and it works), ALK (Total protein) etc...I think these ones don´t just work and I want to find out whether anybody has worked with those I mentionned (with the specification of the phosphorylated sides) and got good data so that they can indicate the name of the company for, most of my antibody are also from cell signalling.
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