Anti-CBX4 Immunofluorescence

June 05, 2014

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Biological Science
Florida State University
Assistant Professor

Company:

Abgent

Product Name:

CBX4 Antibody (C-term)

Catalog Number:

AP2514b

The purpose of this experiment was to identify an antibody that was suitable for detection of CBX4 in human cells by indirect immunofluorescence.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Human 293T 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 Rhodamine 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 relatively well, detecting a nuclear protein with a speckled appearance characteristic of polycomb group proteins

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

No drawbacks were evident with this antibody

The Bottom Line

This antibody is fairly good for immunofluorescence. Abgent provides 100ul of antibody solution which is sufficient for numerous staining experiments.

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