Novus Biologicals
HIF1a antibody
NB100-449
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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.
Immunofluorescence
Primary bone marrow derived macrophages (murine)
1:100 dilution, overnight at 4 deg on a shaker
5% normal goat serum with 0.1% triton X 100
Alexa fluor 647, 1:250 dilution, 2 h, room temperature on a shaker
NA
Confocal imaging
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.
None
Reliable and reproducible
Would be great if it was a fluorescently tagged antibody to eliminate the secondary antibody step
Antibody works in primary mouse macrophages for immunocytochemistry applications.