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400080 Anti-Hypoxia-Inducible Factor 1α Mouse mAb (H1α67)

Anti-Hypoxia-Inducible Factor 1α Mouse mAb (H1α67) MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: Anti-HIF-1α

400080
  
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Overview

Key Spec Table

Host
M
Description
Overview

This product has been discontinued.





Recognizes several proteins of ~120 kDa representing multiple post-translationally modified forms of HIF-1α in induced tissues and cells.
Catalogue Number400080
Brand Family Calbiochem®
SynonymsAnti-HIF-1α
Application Data
Detection of monkey hypoxia-inducible factor 1α by immunoblotting. Samples: Extracts from Cobalt chloride induced COS-7 nuclear (lane 2). Primary antibody: Anti-Hypoxia-Inducible Factor 1α Mouse mAb (H1α67) (Cat. No. 400080) (1:500). Detection: chemiluminescence.

Detection of human hypoxia-inducible factor 1α by immunohistochemistry. Sample: Human glioblastoma multiforme. Primary antibody: Anti-Hypoxia-Inducible Factor 1α Mouse mAb (H1α67) (Cat. No. 400080) (1:100). Detection: fluorescence.
References
ReferencesDuan, et al. 2005. Circulation 111, 2227.
Feldser, D., et al. 1999. Cancer Res. 59, 3915.
Kaelin, W.G. 1999. Nature 399, 203.
Nguyen, S.V., and Claycomb, W.C. 1999. Biochem. Biophys. Res. Comm. 265, 382.5.
Iyer, N.V., et al. 1998. Genes and Development 12, 149.
Semenza, G.L. 1998. J. Lab. Clin. Med. 131, 207.
Product Information
FormLiquid
FormulationIn 150 mM NaCl, 20 mM sodium phosphate.
Preservative≤0.1% sodium azide
Applications
Key Applications Immunoblotting (Western Blotting)
Immunoprecipitation
Paraffin Sections
Application NotesImmunoblotting (1:500-1:1000)
Immunoprecipitation (see comments)
Paraffin Sections (1:100, heat pre-treatment required)
Application CommentsAlso reported to immunoprecipitate HIF-1α from nuclear extracts. Has also been reported to stain formalin-fixed, paraffin-embedded tissues; antigen unmasking using heat and citrate buffer, pH 6.0, is recommended prior to detection of HIF-1α in tissue samples.

Regarding immunoblotting to detect HIF-1α:

HIF-1 alpha is the most rapidly degraded protein ever studied. Upon cellular re-oxygenation it is completely degraded in less than 1 min. It is critical to prep only a few plates/dishes/flasks of cells at a time and to immediately get the cells into ice cold buffers and perform the entire protein prep on ice. HIF-1α is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5% or with treatment using certain agents (CoCl2, DFO, etc.) so proper sample preparation is critical. Upon stabilization HIF-1α translocates to the nucleus. The cleanest immunoblots are always done using nuclear extracts. It is possible to detect HIF-1α in whole cell extracts, but they tend to be much dirtier and the staining is much weaker. Finally, we recommend that positive and negative controls always be run side by side so that it is possible to discern which band is upregulated in the hypoxic sample. Unprocessed HIF-1α is ~95 kDa while the fully post-translationally modified form is ~116 kDa or larger. Additionally, HIF-1α may form a heterodimer with HIF-1β (Duan, et al.).
Depending on the sample, treatment methods, etc. HIF-1α may appear as a single band or as a doublet.

Variables associated with assay conditions will dictate the optimal dilution.
Biological Information
Immunogena recombinant protein consisting of amino acids 432-528 of human HIF-1α fused to GST
ImmunogenHuman
CloneH1α67
HostMouse
IsotypeIgG2b
Concentration Label Please refer to vial label for lot-specific concentration
Storage and Shipping Information
Ship Code Blue Ice Only
Toxicity Standard Handling
Storage +2°C to +8°C
Do not freeze Yes
Global Trade Item Number
Catalogue Number GTIN
400080 0