Anti-H3K4me2 Immunofluorescence

December 27, 2013

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Biological Science
Florida State University
Assistant Professor

Company:

Active Motif

Product Name:

Histone H3 dimethyl Lys4 Rabbit pAb

Catalog Number:

39141

The purpose of this experiment was to identify an antibody that was suitable for detection of histone H3 dimethylated at lysine 4 in human cells by indirect immunofluorescence.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Human hTERT-RPE1 cells

Primary Incubation

1:400 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 Alexa-488 Goat anti-rabbit 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 goat anti-rabbit antibodies

Results Summary

The antibody stains well and detects a pattern of distribution expected for this histone modification.

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

The signal was very strong and as expected for the distribution of this histone modification

The Bad

No drawbacks were evident with this antibody

The Bottom Line

This antibody is good for immunofluorescence. Provision of 200ul ensures countless staining experiments can be performed.

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