Works Reasonably Well for Flow Cytometry

Chemical/Biomedical Engineering
University of Toronto
PhD Candidate

Overall

Quality of Results

Ease-of-Optimization

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Company:

Fischer

Product Name:

DNMT3A

Catalog Number:

PIMA516171

Clone Number:

64B1446

Info: Supplier Page View Product Specs

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An uncojugated antibody that works well for detecting human and mouse DNMT3A once conjugated to biotin.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse splenocytes

Primary Incubation

Fc Block @ 1:100, 30 minutes on ice; fix and permeabilize 20 minutes, 4C

Blocking Agent

2.4G2

Secondary Incubation

Add Biotinylated antibody @ 1:200, 30 min RT

Tertiary Incubation

Add SA-APC @ 1:1600, 30 min RT

Detection

Flow Cytometry (Attune)

Results Summary

Good detection of DNMT3A in total splenocytes above SA-alone stained background. Some populations of cells do not have total separation of DNMT3A+ and DNMT3A- populations, but there is separation between the negative population and high-expressing cells.

DOI or PMID #

N/A

Additional Notes

Unlabeled antibody was biotin conjugated using abcam lightning link type A kit prior to staining

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Summary

The Good

Reasonable staining for flow cytometry

The Bad

Requires conjugation for use in flow cytometry

The Bottom Line

A good antibody for this application since no pre-conjugated antibodies are available

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