Western Blot Using Caspase-3 (3G2) Mouse mAb from Cell Signaling Technology

Klin. Pharmakol. und Tox.
Charite-Berlin
PhD

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Company:

Cell Signaling Technology

Product Name:

Caspase-3 (3G2) Mouse mAb

Catalog Number:

9668S

We performed protein expression analyses in human podocytes regarding the generation of different factors involved in apoptosis regulation, signaling, cell structure maintenance and cell viability. Specifically phospho-specific antibodies were used to analyse the signaling activity normalized againt total protein expression (see Akt, AMPK and mTOR). With this antibody, we performed Western blot analyses of Caspase-3 in cell lysates of human podocytes using the Caspase-3 (3G2) mouse mAb from Cell Signaling Technology.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Cell lysate of human podocytes

Primary Incubation

Dilution 1:500, 16 hours, 4°C

Blocking Agent

5% Skim milk in TBST buffer

Secondary Incubation

Dilution: 1:5000, 1 hour, 24°C

Tertiary Incubation

ECL - 5 min, room temperature

Detection

ECL

Results Summary

A single band was detected after 5 min (Caspase3-band at 35 kDa; see figure).

Additional Notes

None

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Summary

The Good

A single band was detected after 5 min (Caspase3-band at 35 kDa; see figure).

The Bad

No cleaved Caspase 3 was detected in our experiments

The Bottom Line

This antibody is ok for Western blot analyses of total caspase 3 in human podocytes

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