Filter: AllWBIPIHCIF/ICC

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).

Featured Product
KD/KO Validated

HIF-1 alpha Polyclonal antibody

Name: HIF-1 alpha Polyclonal antibody
Brand: Proteintech
SKU: 20960-1-AP
Rating: 3.40 (5 reviews)

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

Cat No. 20960-1-AP

Publications(725)

Reviews (5)

Host / Isotype

Rabbit / IgG

Reactivity

human and More (3)

Applications

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

Synonyms

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

Formulation: PBS and Azide

PBS and Azide

Conjugate: Unconjugated

Unconjugated

CoraLite® Plus 647 FITC Plus

Size/Concentration:

-/ -

Freight/Packing: -

Quantity

- +

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Overview

Tested Applications
Product Information
Publications (725)
Protocols
Reviews (5)

Print datasheet

MSDS SDS

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

Application	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 54 publications below
WB	See 597 publications below
IHC	See 157 publications below
IF	See 107 publications below
IP	See 17 publications below
CoIP	See 16 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 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 Predict reactive species
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
UNIPROT ID	Q16665
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). .

Protocols

Product Specific Protocols
WB protocol for HIF-1 alpha antibody 20960-1-AP	Download protocol
IHC protocol for HIF-1 alpha antibody 20960-1-AP	Download protocol
IF protocol for HIF-1 alpha antibody 20960-1-AP	Download protocol
IP protocol for HIF-1 alpha antibody 20960-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

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 View Article
human	WB	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
human	WB	Mol Cell The mitochondrial DNAJC co-chaperone TCAIM reduces α-ketoglutarate dehydrogenase protein levels to regulate metabolism Authors - Wang Jiahui View Article

Reviews

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