Highly Efficient Human TFAM Antibody

Houston methodist hospital
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

TFAM (D5C8) Rabbit mAb

Catalog Number:

8076S

Clone Number:

D5C8

Info:Supplier Page View Product Specs

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The antibody sensitivity for Western blot analysis is exceptionally high in human cells, including mesenchymal stem cells, HUVECs, and some cancer cell lines. To detect the presence of TFAM protein, the antibody was diluted to 1:1000 in a 5% BSA-containing TBST solution and incubated overnight. The following day, the membrane was incubated with an HRP-conjugated rabbit antibody for 1 hour. The development of HRP resulted in the detection of the TFAM band just 20 seconds later, indicating the effectiveness and speed of the antibody in identifying the target protein.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Human cell (MSC, HUVEC)

Primary Incubation

1/1000 in 5% BSA/TBST for overnight

Blocking Agent

5% BSA

Secondary Incubation

1/10000 HRP-Rabbit IgG for 1 hr in room temperature

Tertiary Incubation

N/A

Detection

HRP for 20 second exposure

Results Summary

To investigate TFAM expression, I used human cells such as MSC, SMC, HUVEC. After inducing TFAM expression, we purified total protein using a protease/phosphatase inhibitor cocktail contained in RIPA buffer. We loaded 20 μg of the purified protein onto an SDS-PAGE gel, and were able to detect the protein band of interest without any background interference. This successful detection of TFAM protein suggests the efficacy of our experimental approach.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Highly efficient and no background.

The Bad

Too expensive.

The Bottom Line

Highly recommend this antibody to the researchers who has an interest to mitochondrial DNA replication study.

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