Medag Is Not Detected in Positive Control

October 09, 2019

Biochemistry
Memorial University
Postdoctoral Researcher

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Company:

Thermo Fisher Scientific

Product Name:

Medag polyclonal antibody

Catalog Number:

PA5-70956

Clone Number:

Polyclonal

I used the Medag antibody to detect the protein in 3T3-L1 cells in different stages of adipogenesis while knocking down the regulated gene CD24.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Mouse, 3T3-L1 cells

Primary Incubation

1:250 in 5% BSA in 1 X TBST for overnight at 4 degrees

Blocking Agent

5% skimmed milk

Secondary Incubation

1:2000 in 5% BSA in 1 X TBST for 1 hr at room temp

Tertiary Incubation

N/A

Detection

Chemiluminiscence

Results Summary

In an attempt to more fully validate the genes that were found to be regulated by CD24 in both 3T3-L1 and primary cells, we attempted to validate at the protein level by western blot. Antibodies for MEDAG (ThermoFisher PA5-70956) is commercially available, however, we were unable to detect the protein with the antibody, even when tested with cell lysates known to express the protein; most likely because of the low quality of the antibody.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

N/A

The Bad

It doesn't detect the Medag protein

The Bottom Line

Please do not buy the product since it is not working in the positive controls as mentioned by the manufacturer. It consumes time and energy to validate the antibody only to know that it is not working.

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