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  • Featured Product
  • KD/KO Validated

HIF-1 alpha Polyclonal antibody

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

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

Validation Data Gallery

Filter:
  • Western Blot (WB) analysis of various lysates using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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.

  • Western Blot (WB) analysis of HeLa cells using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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.

  • Western Blot (WB) analysis of various lysates using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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.

  • Western Blot (WB) analysis of various lysates using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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.

  • Immunoprecipitation (IP) experiment of HeLa cells using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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.

  • Immunohistochemistry (IHC) staining of human thyroid cancer tissue using HIF-1 alpha Polyclonal antibody (20960-1-AP)

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

    Immunohistochemical analysis of paraffin-embedded human thyroid cancer tissue slide using 20960-1-AP (HIF1a antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemistry (IHC) staining of human thyroid cancer tissue using HIF-1 alpha Polyclonal antibody (20960-1-AP)

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

    Immunohistochemical analysis of paraffin-embedded human thyroid cancer tissue slide using 20960-1-AP (HIF1a antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

  • Immunohistochemistry (IHC) staining of human heart tissue using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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).

  • Immunohistochemistry (IHC) staining of human heart tissue using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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).

  • Immunohistochemistry (IHC) staining of human kidney tissue using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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).

  • Immunohistochemistry (IHC) staining of human kidney tissue using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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).

  • Immunohistochemistry (IHC) staining of human thyroid cancer tissue using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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).

  • Immunohistochemistry (IHC) staining of human thyroid cancer tissue using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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).

  • Immunofluorescence (IF) / fluorescent staining of HeLa cells using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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 Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

  • Immunofluorescence (IF) / fluorescent staining of HepG2 cells using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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 Goat Anti-Rabbit IgG(H+L).

  • Immunofluorescence (IF) / fluorescent staining of HepG2 cells using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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 Goat Anti-Rabbit IgG(H+L).

  • Immunofluorescence (IF) / fluorescent staining of HeLa cells using HIF-1 alpha Polyclonal antibody (20960-1-AP)

    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 Goat Anti-Rabbit IgG(H+L), CL594-phalloidin (red).

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Tested Applications

Positive WB detected inCobalt Chloride treated HeLa cells, HeLa cells
Positive IP detected inHeLa cells
Positive IHC detected inhuman 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 inCobalt Chloride treated HeLa cells, Cobalt Chloride treated HepG2 cells

Recommended dilution

ApplicationDilution
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)/ICCIF/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

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 Reactivityhuman, pig, chicken, goat
Host / Isotype Rabbit / IgG
Class Polyclonal
Type Antibody
Immunogen HIF-1 alpha fusion protein Ag15198 相同性解析による交差性が予測される生物種
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 numberBC012527
Gene symbol HIF1A
Gene ID (NCBI) 3091
RRIDAB_10732601
Conjugate Unconjugated
Form Liquid
Purification MethodAntigen affinity purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

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). .

Protocols

Product Specific Protocols
WB protocol for HIF-1 alpha antibody 20960-1-APDownload protocol
IHC protocol for HIF-1 alpha antibody 20960-1-APDownload protocol
IF protocol for HIF-1 alpha antibody 20960-1-APDownload protocol
IP protocol for HIF-1 alpha antibody 20960-1-APDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
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
View Article
humanWB

Cancer Cell

Transcriptional Regulation of the Warburg Effect in Cancer by SIX1.

Authors - Ling Li
View Article
IF

Cancer Cell

Cancer cell autophagy, reprogrammed macrophages, and remodeled vasculature in glioblastoma triggers tumor immunity

Authors - Agnieszka Chryplewicz
View Article

Nat Cell Biol

A single-cell transcriptomic landscape of the lungs of patients with COVID-19.

Authors - Si Wang
View Article
WB

ACS Nano

Augmenting Intracellular Cargo Delivery of Extracellular Vesicles in Hypoxic Tissues through Inhibiting Hypoxia-Induced Endocytic Recycling

Authors - Bide Tong
View Article
IF

Adv Sci (Weinh)

Inflammatory Fibroblast-Like Synoviocyte-Derived Exosomes Aggravate Osteoarthritis via Enhancing Macrophage Glycolysis

Authors - Bin Liu
View Article

Reviews

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

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