Anti-DNMT3B Immunofluorescence

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Biological Science
Florida State University
Assistant Professor

Company:

Santa Cruz Biotechnology, Inc.

Product Name:

Anti-DNMT3B (N-19)

Catalog Number:

sc-10235

The purpose of this experiment was to identify an antibody that was suitable for detection of DNMT3B in human cells by indirect immunofluorescence.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Human hTERT-RPE1 cells

Primary Incubation

1:200 at room temperature for 60 minutes. Staining was performed in 1 x PBS and 0.1% Tween 20

Blocking Agent

Slide was pre blocked with 3% BSA in 1 x PBS and 0.1% Tween 20

Secondary Incubation

1:100 dilution of FITC Donkey anti-Goat secondary diluted in 1 x PBS and 0.1% Tween-20 for 60 minutes at room temperature

Tertiary Incubation

No tertiary incubation was performed

Detection

Direct labeled Donkey anti-Goat antibodies

Results Summary

This antibody detects a nuclear protein. Background is a little high, but does not prevent detecting the nucleus.

Additional Notes

Cells were grown directly on slides and washed briefly with 1 x PBS before fixing and extracting for 10 minutes at room temperature in 4% formaldehyde, 1 x PBS and 0.1% Triton-X100

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Summary

The Good

Good nuclear signal but with some background

The Bad

Background is a little high.

The Bottom Line

This antibody is suitable for detection of DNMT3B by immunofluorescence.

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