Dear all,
I have problem to detect tau (pan and phosphorylated) protein expression, using Western Błot. I've tried increasing the amount od protein (up to 175 ug) and concentration of Ab (Cell Signaling, 1:600) but without succes. The Ponceu stainibg after transfer looks really nice (photo attached). Is anyone have similar problem?
Hi Bartosz,
What antibodies are you using and how long is your primary and secondary incubation for each antibody? These are all factors that could be contributing to your lack of signal
Hi Trevor Price ,
I've tried with Phospho-Tau Family Antibody Sampler Kit #96628 (Cell Sign.) without succes.
Incubation time:
Primary Ab - 12 -16h., overnight
Secondary: 1h, RT, dillution: even the 1:3000 was not efficient.
Wet transfer, overnight, 35V in 4°C.
I prefer blocking with 1% BSA in TBST, not milk.
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