Weak Staining But Works Well Enough

Chemical/Biomedical Engineering
University of Toronto
PhD Candidate

Overall

Quality of Results

Ease-of-Optimization

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Company:

abcam

Product Name:

DNMT3L

Catalog Number:

ab93613

Clone Number:

N117/9

This antibody is acceptable for use in flow cytometry after biotin conjugation but does not perform exceptionally well.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse splenocytes

Primary Incubation

Fc block @ 1:100, 30 minutes RT; Fix/perm 20 minutes 4C

Blocking Agent

2.4G2

Secondary Incubation

Add biotinylated antibody 1:100, 30 minutes RT

Tertiary Incubation

Add SA-APC 1:1600, 30 minutes RT

Detection

Flow cytometry (Attune)

Results Summary

After biotinylation, DNMT3L expressing cells are detectable by flow cytometry with SA-APC, although there is a continuous range of intermediate to highly stained cells.

DOI or PMID #

Additional Notes

Unlabeled antibody was conjugated to biotin using abeam's lightning link type A kit before staining

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Summary

The Good

One of the few commercially available DNMT3L antibodies

The Bad

Not extremely clean staining

The Bottom Line

Reasonable for use flow cytometry although somewhat dirty and expensive for its results

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