Strong Signal: Anti-SAP30 Immunofluorescence

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Biological Science
Florida State University
Assistant Professor

Company:

EMD Millipore

Product Name:

Anti-SAP30

Catalog Number:

06-875

The purpose of this experiment was to identify an antibody that was suitable for detection of SAP30 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 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 detects a nuclear protein with the expected distribution for SAP30 with little to no background.

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 strong and as expected for the distribution of this protein

The Bad

No drawbacks were evident with this antibody

The Bottom Line

This is a good antibody for immunofluorescence.

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