Great Zap-70 Antibody for Western Blotting Using Jurkat Cells

Medicine
Harvard Medical School
Post-doctoral fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Zap-70 antibody

Catalog Number:

3165S

Clone Number:

D1C10E

I study the mechanisms and underlying pathways of asthma development and exacerbations. Here, I determined the levels of total ZAP-70 protein which is a T cell activation marker.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Jurkat T cell lines

Primary Incubation

1:2000 dilution, 18 hours at 4 degrees with gentle shaking

Blocking Agent

5% non-fat dry milk in Tris buffer containing 0.1% Tween-20

Secondary Incubation

HRP-conjugated anti-rabbit antibody 1:2000 dilution, 1 hour at room temperature with gentle shaking

Tertiary Incubation

Not applicable

Detection

ECL based detection

Results Summary

Data showed that ZAP70 protein was present in both samples albeit higher in sample 2.

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Summary

The Good

Specificity

The Bad

Background

The Bottom Line

Overall, it's a good antibody for Western blotting purposes. Blocking needs to be more stringent to achieve best quality data.

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