HIF1a in Immunocytochemistry

Internal medicine
UNIVERSITY OF CINCINNATI
Postdoctoral fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Novus Biologicals

Product Name:

HIF1a antibody

Catalog Number:

NB100-449

The goal was to detect HIF1a localization in primary bone marrow derived macrophages. This product was selected as NB100-449 has been cited in literature for HIF1a detection by western blot.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Primary bone marrow derived macrophages (murine)

Primary Incubation

1:100 dilution, overnight at 4 deg on a shaker

Blocking Agent

5% normal goat serum with 0.1% triton X 100

Secondary Incubation

Alexa fluor 647, 1:250 dilution, 2 h, room temperature on a shaker

Tertiary Incubation

NA

Detection

Confocal imaging

Results Summary

Image (top right) derived from merge of DAPI and AF647 staining. White arrows point at cells containing nuclear HIF1a. Unstained and secondary antibody controls shown in bottom panels, Magnification, 63X. HIF1a nuclear localization was detected in the macrophages as seen in the merge images.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Reliable and reproducible

The Bad

Would be great if it was a fluorescently tagged antibody to eliminate the secondary antibody step

The Bottom Line

Antibody works in primary mouse macrophages for immunocytochemistry applications.

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