Question:

I ran a western of HCMV AD169 infected human foreskin fibroblasts and probed the blot for pp65 using your antibody CA003-100 at a 1:10000 dilution in PBS + .1% Tween 20 + 1% BSA. I didn't see anything on the blot.

I used a different pp65 antibody from one of our collaborators and was able to detect pp65 from the same sample.

The product sheet says a 1:16000 dilution is sufficient to detedt the pp65 protein. Any suggestions as to why I am not detecting pp65 with your antibody? I am setting up some moe blots today and I intend to test a couple different dilutions of your antibody against pp65.

Resolution:

There are many possible reasons for the results you obtained. In order for you to duplicate our results, you need to duplicate our conditions.

Our CMV extract (CV001) is loaded at 10 µg/cm, transferred to BioRad PVDF, probed with the antibody at various dilutions, detected with anti-mouse IgG-AP (Jackson Immunoresearch 1:5,000 of 1 mg/ml) and BCIP/NBT according to the western procedure on our website.

Sounds as if you have the antigen controlled for by way of the other antibody - you may just need to adjust the dilution of our antibody to be appropriate for your assay system.

Hope this helps - let us know how you do with it.