Hard to Detect FIP200 When Small Amount of Lysates Loaded

Cell Biology
Harvard Medical School
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

FIP200 (D10D11) Rabbit mAb

Catalog Number:

12436

I work on selective forms of autophagy, especially focusing on ribophagy and ER-phagy. I have used large numbers of antibodies for proteins related to autophagy. For this FIP200 antibody, KO clones were validated; however, it was not easy to detect FIP200, as the band was too weak.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HCT116 cell lysate

Primary Incubation

1:1000 in 5% BSA in TBST, overnight incubation at 4 degree

Blocking Agent

5% non-fat milk in TBST

Secondary Incubation

1:20000 Licor IRdye800 rabbit

Tertiary Incubation

Washed with TBST three times, 5 min each

Detection

Odyssey clx

Results Summary

I used this antibody to screen FIP200 KO. Since the signal was quite weak, I had to load great amount of the lysate in order to validate clear knock-out.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Okay selectivity

The Bad

Low sensitivity

The Bottom Line

I would try another antibody first before trying this one.

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