Good Antibody for Looking at Insulin Signaling

March 22, 2019

Oncology
Johns Hopkins University
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) Antibody

Catalog Number:

3021

My research focus is on understanding the role of the immune system in regulating blood glucose and adipose tissue. For this experiment, HEK293T cells were serum starved and stimulated with insulin for increasing duration's. Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) signaling was interrogated.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293T cells

Primary Incubation

4% BSA in TBST overnight at 4°C

Blocking Agent

5% milk in TBST

Secondary Incubation

5% milk in TBST for 2 hours at room temperature

Tertiary Incubation

N/A

Detection

ECL

Results Summary

HEK293T cells were serum starved and stimulated with insulin for increasing durations. Phospho-IGF-I Receptor β (Tyr1131)/Insulin Receptor β (Tyr1146) signaling was interrogated.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Antibody gives a nice clean signal

The Bad

Antibody does not work as well when reused

The Bottom Line

The antibody gives a nice clean signal for insulin signaling. However, when the antibody is reused, the signal is much weaker. Usually I need to make up the antibody fresh which increases cost.

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