Good iNOS Antibody from Proteintech

September 27, 2019

ICGS
University of Birmingham
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Proteintech

Product Name:

iNOS

Catalog Number:

18985-1-AP

Clone Number:

Polyclonal

We tried this antibody with 2 cell lines and it worked just fine. However, with a third cell line, there was no band coming up at 105 kDa, only a band at around 70kDA, which might have been non-specific binding. We will use this in the future for other cell lines as well. The antibody produced strong bands at the suggested concentration and we diluted it further.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa cells, human primary cells, hPSCs

Primary Incubation

Standard dilution we use is 1:1000 overnight at 4 degrees

Blocking Agent

5% BSA buffer

Secondary Incubation

1:4000, RT, 60 mins

Tertiary Incubation

Not required

Detection

Immunoblotting

Results Summary

The antibody was only problematic with one cell line, but worked fine for the rest. In the problematic cell line, no band appeared in both control and treated samples (treated should have produced a strong band). There was a non-specific binding at 70 kDa, that was detected several times in different biological replicas, which was weird.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Produced strong bands

The Bad

Did not work properly in all cell lines

The Bottom Line

Proteintech does usually have good antibodies but this one did not produce results as expected. Not sure whether to blame it on the sales representative that recommended it for western blotting, as I looked on the website afterward, and western blotting was not one of the suggested uses for this antibody.

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