Very Strong Signal: Anti-CAF-1 IF

April 22, 2014

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Biological Science
Florida State University
Assistant Professor

Company:

Santa Cruz Biotechnology Inc.

Product Name:

CAF-1 p150 (H-300)

Catalog Number:

sc-10772

The purpose of this experiment was to identify an antibody that was suitable for detection of the chromatin assembly factor CAF-1 in human cells during S-phase 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 stains well and detects a pattern of distribution expected for replication patterns in S-phase.

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

Image Gallery

Summary

The Good

The signal was very strong and as expected for cells in S-phase

The Bad

No drawbacks were evident with this antibody

The Bottom Line

This antibody is good for immunofluorescence. Provision of 1.0ml ensures countless staining experiments can be performed.

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