HIF-1 alpha Polyclonal antibody

• Featured Product
• KD/KO Validated

HIF-1 alpha Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human and More (3)

Applications

WB, IHC, IF/ICC, IP, CoIP, ChIP, RIP, ELISA, Cell treatment

Conjugate

Unconjugated

Publications

654

Reviews (5)

Cat no : 20960-1-AP

Synonyms

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

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Validation Data Gallery

• 

  WB analysis using 20960-1-AP

  Untreated and Cobalt Chloride treated HeLa cells were subjected to SDS PAGE followed by western blot with 20960-1-AP (HIF1a antibody) at dilution of 1:6000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 20960-1-AP

  WB result of HIF-1 alpha antibody (20960-1-AP; 1:6000; 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.

• 

  WB analysis using 20960-1-AP

  Untreated and Cobalt Chloride treated HeLa cells were subjected to SDS PAGE followed by western blot with 20960-1-AP (HIF1a antibody) at dilution of 1:6000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 20960-1-AP

  Untreated and Cobalt Chloride treated HeLa cells were subjected to SDS PAGE followed by western blot with 20960-1-AP (HIF1a antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of HeLa using 20960-1-AP

  IP result of anti-HIF-1 alpha (IP:20960-1-AP, 4ug; Detection:20960-1-AP 1:300) with HeLa cells lysate 4000ug.

• 

  IHC staining of human thyroid cancer using 20960-1-AP

  Immunohistochemical analysis of paraffin-embedded human thyroid cancer tissue slide using 20960-1-AP (HIF-1 alpha antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human thyroid cancer using 20960-1-AP

  Immunohistochemical analysis of paraffin-embedded human thyroid cancer tissue slide using 20960-1-AP (HIF-1 alpha antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human heart using 20960-1-AP

  Immunohistochemical analysis of paraffin-embedded human heart using 20960-1-AP (HIF1A antibody) at dilution of 1:100 (under 10x lens).

• 

  IHC staining of human heart using 20960-1-AP

  Immunohistochemical analysis of paraffin-embedded human heart using 20960-1-AP (HIF1A antibody) at dilution of 1:100 (under 40x lens).

• 

  IHC staining of human kidney using 20960-1-AP

  Immunohistochemical analysis of paraffin-embedded human kidney using 20960-1-AP (HIF1A antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human kidney using 20960-1-AP

  Immunohistochemical analysis of paraffin-embedded human kidney using 20960-1-AP (HIF1A antibody) at dilution of 1:50 (under 40x lens).

• 

  IHC staining of human thyroid cancer using 20960-1-AP

  Immunohistochemical analysis of paraffin-embedded human thyroid cancer using 20960-1-AP (HIF1A antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human thyroid cancer using 20960-1-AP

  Immunohistochemical analysis of paraffin-embedded human thyroid cancer using 20960-1-AP (HIF1A antibody) at dilution of 1:50 (under 40x lens).

• 

  IF Staining of HeLa using 20960-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed Cobalt Chloride treated HeLa cells using HIF-1 alpha antibody (20960-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

• 

  IF Staining of HepG2 using 20960-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed Cobalt Chloride treated HepG2 cells using HIF1a antibody (20960-1-AP) at dilution of 1:1800 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HepG2 using 20960-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed Cobalt Chloride treated HepG2 cells using HIF1a antibody (20960-1-AP) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of HeLa using 20960-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed Cobalt Chloride treated HeLa cells using HIF-1 alpha antibody (20960-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

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Tested Applications

• Positive WB detected in: Cobalt Chloride treated HeLa cells, HeLa cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human thyroid cancer tissue, human heart tissue, human kidney 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

Samples need to be treated with hypoxia stimulation.

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:12000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• 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.

Published Applications

• KD/KO: See 50 publications below
• WB: See 545 publications below
• IHC: See 140 publications below
• IF: See 93 publications below
• IP: See 17 publications below
• CoIP: See 12 publications below
• ChIP: See 17 publications below
• RIP: See 1 publications below

Product Information

20960-1-AP targets HIF-1 alpha in WB, IHC, IF/ICC, IP, CoIP, ChIP, RIP, ELISA, Cell treatment applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, pig, chicken, goat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• 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_10732601
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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). .

Publications

Species	Application	Title
	WB	Gastroenterology; Pancreatic acinar cells-derived sphingosine-1-phosphate contributes to fibrosis of chronic pancreatitis via inducing autophagy and activation of pancreatic stellate cells; Authors - Decai Wang
human	WB	Cancer Cell; Transcriptional Regulation of the Warburg Effect in Cancer by SIX1.; Authors - Ling Li
	IF	Cancer Cell; Cancer cell autophagy, reprogrammed macrophages, and remodeled vasculature in glioblastoma triggers tumor immunity; Authors - Agnieszka Chryplewicz
		Nat Cell Biol; A single-cell transcriptomic landscape of the lungs of patients with COVID-19.; Authors - Si Wang
	WB	ACS Nano; Augmenting Intracellular Cargo Delivery of Extracellular Vesicles in Hypoxic Tissues through Inhibiting Hypoxia-Induced Endocytic Recycling; Authors - Bide Tong
	IF	Adv Sci (Weinh); Inflammatory Fibroblast-Like Synoviocyte-Derived Exosomes Aggravate Osteoarthritis via Enhancing Macrophage Glycolysis; Authors - Bin Liu

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Carlo (Verified Customer) (01-18-2023)

The quality of the band in western blot is well defined

• Applications: Western Blot
• Primary Antibody Dilution: 1:200
• Cell Tissue Type: Neuroblastoma cell line

FH

Davide (Verified Customer) (09-20-2022)

The image uploaded refers to a hypoxic condition O2<1%. The antibody signal was aspecific and generated aspecific binding in the middle of the organoid.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:100
• Cell Tissue Type: Cholangiocyte organoids

HIF-1 alpha Antibody Immunofluorescence validation (1:100 dilution) in Cholangiocyte organoids (Cat no:20960-1-AP)

FH

Christian (Verified Customer) (01-07-2022)

Unfortunately much background an many unspecific bands in lysates of human chondrocytes and BV2 microclia cells (less unspecific bands in rat chondrocytes).

• Applications: Western Blot
• Cell Tissue Type: human chondrocytes

HIF-1 alpha Antibody Western Blot validation ( dilution) in human chondrocytes (Cat no:20960-1-AP)

FH

Diane (Verified Customer) (02-17-2021)

Separated on a 7.5% SDS PAGE gel, transferred to nitrocellulose membrane. Block with 5% dry milk/PBST at room temperature for one hour. 1:1000 dilution of the primary antibody in blocking solution. Incubated overnight at 4 degrees C on a rocker platform. Secondary antibody 1:1000 secondary (anti rabbit HRP) incubation for two hours at room temperature. Visualization with the Opti-4CN chromogenic substrate kit (BioRad).

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: primary glioblastoma cell lysates

HIF-1 alpha Antibody Western Blot validation (1:1000 dilution) in primary glioblastoma cell lysates (Cat no:20960-1-AP)

FH

Chun (Verified Customer) (12-05-2019)

A good antibody

• Applications: Western Blot,
• Primary Antibody Dilution: 1:1,000
• Cell Tissue Type: HeLa cells

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