I will be performing Western blotting, to detect HSV-1 infected cell proteins, with your antigen products, H1V016-1 (lot # C0010056) and AV042-1 (lot #116017); and your anti-ICP0, anti-ICP4 and anti-ICP5 antibodies. Your literature indicates a Western Blot titer of 10ug/cm. Does this mean that a loading of 10 ug of protein is recommended for lanes that are 1 cm wide? I will be using a chemiluminescent detection method. I would appreciate any recommendations you might have, as to an initial concentration of each antigen product to test. I will be using BioRad Criterion gels, approximate lane width, 0.5 cm.
Thank you for your time and attention
Resolution:
You are correct. For a 0.5 cm lane, you would load 5 µg to duplicate our conditions.
Regarding chemiluminescence, this methodology is normally more sensitive than the BCIP/NBT used in our photography. I think that I would go with one of the more dilute concentrations shown on the Certificate of Analysis and use exposure time to make final adjustments if the signal is too strong.
I hope this helps
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