Strong Signal: Anti-BMI1 IF

April 07, 2014

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Biological Science
Florida State University
Assistant Professor

Company:

EMD Millipore

Product Name:

Anti-Bmi-1, clone F6

Catalog Number:

05-637

The purpose of this experiment was to identify an antibody that was suitable for detection of BMI1 in human cells by indirect immunofluorescence.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Human 293T 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 Rhodamine Goat anti-mouse 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-mouse antibodies

Results Summary

The mouse monoclonal antibody stains well showing the expected nuclear distribution of BMI1. The intense foci shown in the image correspond to the human heterochromatic inactive X chromosome. 293T are polyploid and can have anywhere from 0 - 4 inactive X chromosome signals per nucleus. The image shows the distribution of BMI1 (red) merged with DAPI (blue).

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 is very strong and the staining pattern as anticipated for this protein.

The Bad

No drawbacks were evident with this antibody

The Bottom Line

Very good antibody for immunofluorescence. Millipore provides 100ul, which at 1:400 dilution for staining allows countless experiments.

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