so, we're using HA-taaged receptors for surface ELISA, but unfortunately our antidbody seems to be not specific or our cell got lysed during the procedure? We can detect also HA-tagged intracellulare proteins without permeabilization step in between - a protein thought to be a negativ control. Did u have similar problems? What a antibody do u use? any tips in general? thanks a lot!
HEK-293
Transfection: Lipofectamine
Fixation: 4% Paraformaldehyd @ rt
Block: full media (including FCS)
Antibody: Roche antiHA-POD
Thx a lot,
cheers,
l.
Xianfeng Wang
For surface expression, maybe fixing cells with 1% formaldehyde (no methanol) in PBS is better.
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