Anti-CHD7 IF

June 09, 2014

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Biological Science
Florida State University
Assistant Professor

Company:

Abcam

Product Name:

Anti-CHD7 Rabbit Polyclonal

Catalog Number:

ab31824

The purpose of this experiment was to identify an antibody that was suitable for detection of CHD7 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 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 detects a nuclear protein, but background in the cytoplasm is relatively high.

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 is relatively strong, but with some background

The Bad

The background makes it hard to image in cells that have low levels of CHD7 expression

The Bottom Line

The antibody is suitable for immunofluorescence. Abcam provides sufficient volume for numerous staining experiments.

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