Western Blot Analyses Using Anti-Phospho-mTOR (Ser2448)

Klin. Pharmakol. und Tox.
Charite-Berlin
PhD

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Company:

Merck Millipore

Product Name:

Anti-phospho-mTOR (Ser2448)

Catalog Number:

09-213

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). Here we performed Western blot analyses of phospho-mTOR in cell lysates of human podocytes using the anti-phospho-mTOR (Ser2448) 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

Several bands were detected after 5 min including a phospho-mTOR band at 289 kDa (see figure).

Additional Notes

Related Categories

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Summary

The Good

phospho-mTOR was detected at 289 kDa

The Bad

several other bands were detected

The Bottom Line

This antibody is ok for Western blot analyses of phospho-mTOR in cell lysates of human podocytes

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