HIF-1 alpha antibody (pAb)

RRID: AB_2614934
Aliases: HIF1A, HIF1, MOP1, PASD8, HIF-1A, bHLHe78, HIF-1alpha, HIF1-ALPHA
Catalog No: 39665 Format: 100 µl ¥88,000 Buy
Catalog No: 39666 Format: 10 µl ¥24,000 Buy

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Antibody Type:
Polyclonal
Isotype:
Serum
Purification:
None
Host:
Rabbit
Molecular Weight:
100 kDa
Reactivity:
Human

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 5 µl per ChIP
ChIP-Seq: 5 µl per ChIP-Seq
WB: 1:500 - 1:1,000 dilution

The addition of 0.1% Tween 20 in the blocking buffer and primary antibody incubation buffer is recommended to aid in detection by Western blot. Individual optimization may be required.

ChIP-validated antibodies are available for this target: Catalog No. 39253.

For HIF-1 alpha, we also offer AbFlex® HIF-1 alpha Recombinant Antibody (rAb). For details, see Catalog No. 91139.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

ChIP-qPCR
WB
IF

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

This HIF-1 alpha antibody was raised against a synthetic peptide corresponding to the C-terminal region of human HIF-1 alpha.

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

 

HIF-1 alpha antibody (pAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 5 µl of HIF-1 alpha antibody and 25 µg of chromatin from hypoxic Huh7 cells. ChIP DNA was sequenced on the Illumina HiSeq and 19 million sequence tags were mapped to identify Hif-1 alpha binding sites across the genome. The images show binding in the promoters of Hif1a target genes that regulate metabolism. 

HIF-1 alpha antibody (pAb) tested by ChIP.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 5 µl of HIF-1 alpha antibody and 25 µg of chromatin from hypoxic Huh7 cells. ChIP DNA was used in qPCR with the control primer pairs or gene-specific primer pairs as indicated. Data are presented as Binding Events Detected per 1000 Cells using Active Motif's Epigenetic Services normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

HIF-1 alpha antibody (pAb) tested by Western blot.
Detection of HIF-1 alpha by Western blot analysis with 20 µg COS-7 nuclear extract either untreated (lane 1) or treated with CoCl2 (lane 2).

Background

Hypoxia-inducible factor-1 (HIF1) is a transcription factor involved in the regulation of oxygen homeostasis. It regulates the physiological responses to low oxygen levels (hypoxia) and the pathophysiology of heart attack, cancer, stroke and chronic lung disease. HIF-1 is a heterodimeric protein that consists of two subunits, HIF-1 alpha and HIF-1 beta (also known as ARNT1, the aryl hydrocarbon nuclear translocator). While HIF-1 beta is constitutively expressed, the expression of HIF-1 alpha is induced by oxygen concentrations below 6%. Heterodimerization is required to bind the hypoxia response element (HRE).

Positive Control

Active Motif's COS-7 (CoCl2 treated) nuclear extract (Catalog No. 40600) can be used as a positive control for HIF-1 alpha.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with HIF-1 alpha antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting