Overly Bright Dclk1 Antibody

Chemical/Biomedical Engineering
University of Toronto
PhD Candidate

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Quality of Results

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Company:

Abcam

Product Name:

Alexa Fluor 647 Anti-DCAMKL1 antibody

Catalog Number:

ab202755

Clone Number:

N/A

Info:Supplier Page View Product Specs

Browse Similar Products: DCAMKL1 Antibodies

We are interested in studying the mechanisms responsible for ensuring T cell self tolerance that rely on thyme and extra-thymic Aire expressing cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse thymus

Primary Incubation

Live dead: Zombie NIR LD 1:4000 in 100ul FACS for 20 min at RT

Blocking Agent

2.4G2

Secondary Incubation

Surface stains: in 100uL FACS for 30 min at 4C

Tertiary Incubation

PFA (15 min RT) + FoxP3 Fix/Perm kit (30 min 4C), then intracellular stain Dclk1 1:3000 in 100uL perm buffer for 1hr at RT

Detection

Cytek Aurora spectral flow cytometry

Results Summary

This antibody is extremely bright by spectral FC, with no detectable negative population of mTECs (despite the majority of mTECs being DCLK1- cells) even with extremely low concentrations of antibody in the staining buffer. Requires significant titration to get to work.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Very bright.

The Bad

Can't detect negative populations with normal staining conditions.

The Bottom Line

Not a good antibody for detecting thyme tuft cells.

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