Western Blot Using Anti-Phospho-AMPK1 (Thr174) (#09-290)

Klin. Pharmakol. und Tox.
Charite-Berlin
PhD

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Company:

Merck Millipore

Product Name:

Anti-phospho-AMPK1 (Thr174)

Catalog Number:

09-290

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). In this experiment we performed Western blot analyses of phosphorylated AMPK1 protein in cell lysates of human Podocytes using the Anti-phospho-AMPK1 (Thr174) (#09-290) antibody

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% milk powder in TBST buffer

Secondary Incubation

Dilution: 1:5000, 1 hour, room temperature

Tertiary Incubation

ECL - 5 min, room temperature

Detection

ECL detection

Results Summary

Two bands including one for phospho-AMPK1 (63 kDa) was detected after 5 min (see fig.)

Additional Notes

None

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Summary

The Good

Phospho-AMPK1 (63 kDa) was detected

The Bad

A second band was detected using this antibody

The Bottom Line

This antibody is okay for Western blot analyses of phospho-AMPK in human podocytes

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