Phospho-S6 (Ser235/236) APC Antibody

August 09, 2017

SNS
University of California, Merced
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (APC Conjugate)

Catalog Number:

14733

My goal is to study certain signaling pathways in populations of T cells in autoimmune disease. A couple of pathways I study involve S6 phosphorylation and since it is far downstream, I've been using it to do a lot of my experimental troubleshooting. This particular antibody has been working very well.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse lymphocytes

Primary Incubation

1:50, 1hour, RT on fixed and permeabilized cells

Blocking Agent

Fc Block or anti-CD16/32 blocking antibody

Secondary Incubation

None

Tertiary Incubation

None

Detection

Fluorescence

Results Summary

Generate histograms of the phospho-S6 fluorescence and compare unstimulated to stimulated cells to determine changes in phosphorylation levels. Using PMA/Ionomycin as a positive control is recommended, as it gives a very distinct peak (see attached photo, red=PMA/ionomycin)

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Clean, consistent results

The Bad

High concentration needed for staining

The Bottom Line

This has been a fantastic antibody for troubleshooting other aspects of my experiments as it works consistently and PMA/ionomycin can be used as a clean positive control. I definitely recommend

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