FLAG WB Antibody

Hi Everyone

Hope all of you are safe. I am wondering if anyone can suggest an antibody to detect 3X FLAG by western. I am currently using Rabbit monoclonal antibody HRP conjugate from cell signaling. I tried with 1:125 dilution but still the band is very faint. Any help will be highly appreciated.

Regards.
Rudra Banerjee
Srayko Lab
U of Alberta, Canada.

Hi Rudra -
My lab uses the M2 mouse monoclonal Ab available through Sigma for western blots and other applications.
Good luck!
Abby

Thank You so much Abby for your valuable reply. I will definitely try that one.

I’ll second Abby’s vote for Sigma M2. Works great for 3xFLAG western blots in our hands. We like the M2-HRP conjugate; makes experiments faster as you can skip the 2º antibody step.

Cheers,

Jordan

Thank You very much Jordan. I am now trying to standardize the concentration of sigma M2. I previously used HRP-conjugate from Cell Signaling but that did not work. I used the Sigma M2 for Immunostaining and it did work perfectly but do not know why it gives so much pain in the WB !