Antibody With Consistent, High Quality Performance in IF and Western Blotting

April 22, 2024

Sanford Burnham Prebys Medical Discovery institute

Overall

Quality of Results

Ease-of-Optimization

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Company:

Sigma Aldrich

Product Name:

Anti-FMR1 Antibody

Catalog Number:

F4055-200UL

Clone Number:

N/A

The study I conducted primarily involved investigating the role of Fragile X mental Retardation Protein (FMRP) in the post-transcriptional regulation of muscle stem cell quiescence. The mentioned antibody was used in the Immunostaining of cells and single muscle fibers from EDL muscle of mouse origin. This was an excellent antibody, with consistent performance in IF and Western Blotting.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse Single muscle fiber (Extensor Digitorum Longus) and C2C12 myoblasts

Primary Incubation

1:200

Blocking Agent

10% Fetal Bovine Serum or 10% Horse Serum in 0.25% Triton X100 PBS

Secondary Incubation

Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488

Tertiary Incubation

NA

Detection

Fluorescence microscope

Results Summary

This antibody was instrumental in finding that quiescence muscle stem cells forms stress granule , of which FMRP is a key component. Fmrp is also a known translation repressor This suggested that quiescent muscle stem cells, which are reduced in translation, potentially store the few transcripts produced (example- Myf5, a muscle lineage specific transcription regulator), but not needed for quiescence maintenance, in biomolecular condensates. Upon stem cell activation they are immediately taken to polysomes for translation, suggesting that these cells are poised to enter skeletal muscle lineage and that Fmrp potentially plays a key role in cell state transition by regulating mRNA utilization.

DOI or PMID #

https://doi.org/10.1186/s13395-021-00270-9

Additional Notes

In the attached image, the green channel is FMRP in muscle stem cells

Related Categories

Image Gallery

Summary

The Good

Consistent, high quality performance. Can be used for both mouse and human samples.

The Bad

Not tested Cross linking Immunoprecipitation (CLIP). FMRP being an RNA binding protein can be pulled down with an antibody against the protein.

The Bottom Line

Antibody is of great quality and can potentially be also used for RNA IP.

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