Difficult to Optimize but Good Antibody

July 10, 2014

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
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Medicine
UCSD
Post-doc

Company:

UCSD

Product Name:

p15/p16 antibody

Catalog Number:

sc-28260

We investigate the human genome and its role in medical applications.

I wanted to use this antibody as a marker for aging caused H-Ras induced senescent in BJ cells. Cells were transfected with H-Ras and grown for 12 days which leads to induction of aging associated marks including p16 protein.

Experimental Design and Results Summary

Applications

Western Blot

Sample

BJ cells transfected with H-Ras vector

Primary Incubation

Dilution-1:800, Incubated at 4 degree evernight in BSA+TBST

Blocking Agent

5% BSA

Secondary Incubation

1:1000 anti-rabbit-HRP for 2 hrs at room temprature

Tertiary Incubation

N/A

Detection

ECL

Results Summary

Since the amount of p16 proteins is very low even after induction I got faint bands at shorter exposure times. But got a relatively strong band upon 10 minutes of exposure.

Additional Notes

None

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Summary

The Good

Limited number of p16 antibodies are available in the market. You get a good amount from Santa Cruz and so you do not have to worry about a new lot.

The Bad

Detection is difficult but mostly because of low abundance of p16 protein

The Bottom Line

Although it took me time to optimize the antibody but I would use this antibody for p16 WB.

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