HIF-1 alpha Recombinant antibody

• Featured Product
• KD/KO Validated

HIF-1 alpha Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, ELISA

Conjugate

Unconjugated

CloneNo.

2K1

Cat no : 80933-1-RR

Synonyms

HIF1A, HIF 1 alpha, Class E basic helix-loop-helix protein 78, Basic-helix-loop-helix-PAS protein MOP1, ARNT-interacting protein

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Validation Data Gallery

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  WB analysis of HeLa using 80933-1-RR

  Untreated and Cobalt Chloride treated HeLa cells were subjected to SDS PAGE followed by western blot with 80933-1-RR (HIF1a antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 80933-1-RR

  WB result of HIF-1 alpha antibody (80933-1-RR; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-HIF-1 alpha transfected HeLa cells. Sample 1: non-treated sh-Control transfected HeLa cells, Sample 2: Cobalt Chloride treated sh-Control transfected HeLa cells, Sample 3: Cobalt Chloride treated sh-HIF-1 alpha transfected HeLa cells.

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  IHC staining of human colon using 80933-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 80933-1-RR (HIF-1 alpha antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human stomach using 80933-1-RR

  Immunohistochemical analysis of paraffin-embedded human stomach tissue slide using 80933-1-RR (HIF-1 alpha antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human thyroid cancer using 80933-1-RR

  Immunohistochemical analysis of paraffin-embedded human thyroid cancer tissue slide using 80933-1-RR (HIF1a antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human thyroid cancer using 80933-1-RR

  Immunohistochemical analysis of paraffin-embedded human thyroid cancer tissue slide using 80933-1-RR (HIF1a antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human stomach cancer using 80933-1-RR

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 80933-1-RR (HIF-1 alpha antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HeLa using 80933-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed Cobalt Chloride treated HeLa cells using HIF-1 alpha antibody (80933-1-RR, Clone: 2K1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

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  IF Staining of HepG2 using 80933-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed Cobalt Chloride treated HepG2 cells using HIF1a antibody (80933-1-RR, Clone: 2K1 ) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), 594+phalloidin (red).

• 

  IF Staining of HepG2 using 80933-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed Cobalt Chloride treated HepG2 cells using HIF1a antibody (80933-1-RR, Clone: 2K1 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  Affinity and Kinetic Characterization of 80933-1-RR

  Biolayer interferometry (BLl) kinetic assays of 80933-1-RR against Human HIF-1 alpha were performed. The affinity constant is below 1 pM.

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Tested Applications

• Positive WB detected in: Cobalt Chloride treated HeLa cells, HeLa cells
• Positive IHC detected in: human thyroid cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: Cobalt Chloride treated HeLa cells, Cobalt Chloride treated HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

80933-1-RR targets HIF-1 alpha in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: HIF-1 alpha fusion protein Ag15198
• Full Name: hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
• Calculated Molecular Weight: 826 aa, 93 kDa
• Observed Molecular Weight: 120 kDa
• GenBank Accession Number: BC012527
• Gene Symbol: HIF1A
• Gene ID (NCBI): 3091
• RRID: AB_2918921
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

HIF1a, the major regulator of the cellular responses to hypoxia, consists of an oxygen-sensitive subunit, HIF1 alpha (HIF1A), and an oxygen-insensitive subunit, HIF1 beta (arylhydrocarbon receptor nuclear transporter [ARNT]). Under normal oxygen conditions, HIF1a is continuously produced and destroyed, in a process involving hydroxylation, interaction with von Hippel-Lindau (VHL) protein, polyubiquitylation and subsequent proteasomal degradation. Under hypoxic conditions, hydroxylation is impaired and HIF1a is stabilized. HIF1a localizes in cytoplasm in normoxia, but it can translocate into nuclear in response to hypoxia. The calculated molecular weight of HIF1a is 93 kDa, but the modified protein HIF1a is about 110-120kDa (PMID: 11698256, PMID: 7539918).