Western Blot with phospho-Tuberin S1387 antibody

Radiation Oncology
Ohio State University
postdoctoral researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Phospho-Tuberin/TSC2 (Ser1387) Antibody

Catalog Number:

5584

This antibody was used to detect levels of phospho-tuberin S1387 in cancer cell lines. When loading 50 ug of cell lysate, we were able to detect a strong signal for phospho-tuberin with this antibody.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Cancer cell line lysates (U87)

Primary Incubation

Overnight in TBST containing 0.5% milk and 1:1000 antibody

Blocking Agent

5% milk in TBST

Secondary Incubation

2 hours in TBST containing 0.5% milk and 1:2000 of HRP-conjugated anti-rabbit secondary from Cell Signaling (#7074)

Tertiary Incubation

NA

Detection

Chemiluminescent detection with Millipore Immobilon Western HRP substrate (cat # WBKLS0500)

Results Summary

Using the high sensitivity chemiluminscent HRP substrate from Millipore, we detected strong signal for phospho-tuberin in several cancer cell lines, including U87.

Additional Notes

None

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Summary

The Good

Antibody worked the first time. No optimization needed.

The Bad

Large amount of cell lysate needed to detect phospho-tuberin (50 ug), but this is true with most phospho-antibodies.

The Bottom Line

Antibody highly recommended

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