Customer Experience Using Novus' PBP Antibody

Mon, 04/23/2012 - 10:05


I first tried the PBP antibody (NB110-93495) in June of 2010 and it worked well. I picked this antibody because it had been tested in rat tissue, so I was confident it would work for my rat samples. I stored the PBP antibody in 20ul aliquots after it arrived then stored it at -20C and used it over a 3 month period. I ran a Western blot with 15ug of a RIPA whole cell lystate from WKPT cells a rat kidney immortalized cell line derived from the S1 proximal tubule segment. I used a 10% SDS-PAGE gel and transferred to PVDF membrane for 1 hour in transfer buffer with 10% methanol. I then blocked with odyssey blocking buffer for 2 hours at RT.

Western Blot: PBP Antibody

I probed with the PBP primary antibody with a 1:1000 dilution over night at 4C. I incubated 1 hour with Licor goat anti rabbit 680nm secondary then imaged on a odyssey scanner. I saw a band at about 22kDa, the blot was clear of any other containment bands. I used this PBP antibody several times with similar results.  I wanted to ultimately quantify the PBP band and checked for saturation of the antibody in my samples by also running a blot loading 5, 10, and 15ug of my sample to check for linearity when diluting the antibody 1:1000. The antibody proved to not be saturated and linear intensity was detected across the three amounts. I have only tried it with WKPT samples but it worked great and I would recommend to others!

This guest blog was submitted by Novus customer, Dana Freund of Colorado State University.

Novus Biologicals offers PBP reagents for your research needs including:

 

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