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

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Synonyme: Anti-HIF-1α

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      Übersicht

      Replacement Information

      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
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Standard Handling
      Storage +2°C to +8°C
      Do not freeze Yes
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Bestellnummer GTIN
      400080 0

      Documentation

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

      TitelChargennummer
      400080

      Literatur

      Übersicht
      Duan, 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.
      Datenblatt

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision05-January-2010 JSW
      SynonymsAnti-HIF-1α
      ApplicationImmunoblotting (1:500-1:1000)
      Immunoprecipitation (see comments)
      Paraffin Sections (1:100, heat pre-treatment required)
      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.
      DescriptionProtein G purified mouse monoclonal antibody. Recognizes several proteins of ~120 kDa representing multiple post-translationally modified forms of HIF-1α.
      BackgroundHIF-1 is a nuclear protein that activates gene transcription in response to reduced cellular O2 concentration. HIF-1 activates the transcription of EPO, VEGF, iNOS, heme oxygenase 1, and several other critical intracellular responses to hypoxia. HIF-1 is a heterodimer composed of 2 basic-helix-loop-helix PAS subunits, HIF-1α and HIF-1β. Both subunits are induced by hypoxia and rapidly decay upon return to normoxia. Recent research suggests the ability to regulate hypoxia-inducible factors may be related to tumor-related angiogenesis in certain cancers.
      HostMouse
      Immunogen speciesHuman
      Immunogena recombinant protein consisting of amino acids 432-528 of human HIF-1α fused to GST
      CloneH1α67
      IsotypeIgG2b
      Speciesbovine, ferret, hamster, human, monkey, mouse, porcine, rabbit, rat, sheep
      FormLiquid
      FormulationIn 150 mM NaCl, 20 mM sodium phosphate.
      Concentration Label Please refer to vial label for lot-specific concentration
      Preservative≤0.1% sodium azide
      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.
      Storage +2°C to +8°C
      Do Not Freeze Yes
      Toxicity Standard Handling
      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.