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HIF1 alpha antibody

KO/KD Orthogonal Overexpression
GTX127309_41234_20170904_WB_O2_shRNA_watermark_18121410_414.jpg
GTX127309_41031_IFA_18121410_467.jpg
GTX127309_42172_20150813_WB_Hypoxia_18121410_921.jpg
GTX127309_41577_IHC_18121410_579.jpg
GTX127309_42662_20170118_IFA_18121410_957.jpg
GTX127309_42123_20151016_IP_18121410_923.jpg
GTX127309_42172_20160128_ChIP_18121410_220.jpg
GTX127309_42123_20150924_IFA_2_18121410_338.jpg
GTX127309_42389_20160728_WB_M_CoCl2_18121410_165.jpg
GTX127309_41577_WB_CoCl2_18121410_886.jpg
GTX127309_42760_20170316_WB_CoCl2_18121410_159.jpg
GTX127309_42389_20160526_WB_B_18121410_315.jpg

Cat No. GTX127309

Host Rabbit
Clonality Polyclonal
Isotype IgG
Application WB, ICC/IF, IHC-P, IHC-Fr, IP, ChIP assay
Reactivity Human, Mouse, Rat, Bovine
APPLICATION

Application Note

*Optimal dilutions/concentrations should be determined by the researcher.
Application Dilution
WB 1:500-1:3000
ICC/IF 1:100-1:1000
IHC-P 1:100-1:1000
IHC-Fr Assay dependent
IP 1:100-1:570
ChIP assay Assay dependent
Not tested in other applications.

Calculated MW

93 kDa. ( Note )

Positive Control

MCF-7 , HeLa(500uM CoCl for 24hr) , NIH-3T3 (500 μM CoCl2 treatment for 24 hr) , HepG2 whole cell lysate/extract (1% O2 treatment for 24hr) , 293T transfected HIF1A , HepG2 ( 200 μM CoCl2 treatment for 24 hr) , HepG2 (500 μM CoCl2 treatment for 24 hr)

Specificity/Sensitivity

Knockdown/Knockout validation was supported by references (PMID:25428606)

Predict Reactivity

Rabbit, Dog, Pig(>80% identity)
PROPERTIES

Form

Liquid

Buffer

1XPBS, 1% BSA, 20% Glycerol (pH7). 0.025% ProClin 300 was added as a preservative.

Storage

Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.

Concentration

0.26mg/ml(Please refer to the vial label for the specific concentration.)

Antigen Species

Human

Immunogen

Recombinant protein encompassing a sequence within the C-terminus region of human HIF1 alpha. The exact sequence is proprietary.

Purification

Purified by antigen-affinity chromatography.

Conjugation

Unconjugated

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET

Synonyms

HIF-1alpha antibody, HIF1 antibody, HIF1-ALPHA antibody, MOP1 antibody, PASD8 antibody, bHLHe78 antibody, HIF1A antibody, basic-helix-loop-helix-PAS protein MOP1 antibody, class E basic helix-loop-helix protein 78 antibody, ARNT-interacting protein antibody, member of PAS superfamily 1 antibody, hypoxia-inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor) antibody, hypoxia-inducible factor 1-alpha antibody, hypoxia-inducible factor 1 alpha isoform I.3 antibody, PAS domain-containing protein 8 antibody, member of PAS protein 1 antibody, HIF-1-alpha antibody, ARNT interacting protein antibody, hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor) antibody

Cellular Localization

Cytoplasm , Nucleus

Background

Hypoxia-inducible factor-1 (HIF1) is a transcription factor found in mammalian cells cultured under reduced oxygen tension that plays an essential role in cellular and systemic homeostatic responses to hypoxia. HIF1 is a heterodimer composed of an alpha subunit and a beta subunit. The beta subunit has been identified as the aryl hydrocarbon receptor nuclear translocator (ARNT). This gene encodes the alpha subunit of HIF-1. Overexpression of a natural antisense transcript (aHIF) of this gene has been shown to be associated with nonpapillary renal carcinomas. Two alternative transcripts encoding different isoforms have been identified. [provided by RefSeq]

Database

Research Area

DATA IMAGES
GTX127309_41234_20170904_WB_O2_shRNA_watermark_18121410_414.jpg

GTX127309 WB Image

KO/KD

Non-transfected (–) and transfected (+) MCF-7 whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with HIF1 alpha antibody (GTX127309) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX127309_41031_IFA_18121410_467.jpg

GTX127309 ICC/IF Image

HIF1 alpha antibody detects HIF1 alpha protein at nucleus in CoCl2-treated samples by confocal immunofluorescent analysis.
Samples: Untreated (left panel) and 500μM CoCl2 treated (right panel) HeLa cells for 24 hrs were fixed in 4% paraformaldehyde for 15 min
Green: HIF1alpha protein stained by HIF1 alpha antibody (GTX127309) diluted at 1:1000
Red: Alpha-tubulin, a cytoskeleton marker, stained by Alpha-tubulin antibody (GTX11304) diluted at 1:2500
Blue: Hoechst 33342 staining.
Scale bar = 10 μm.
Images captured by Olympus FV10i Confocal Laser Scanning Microscope

GTX127309_42172_20150813_WB_Hypoxia_18121410_921.jpg

GTX127309 WB Image

Orthogonal

HIF1 alpha antibody detects HIF1 alpha protein by western blot analysis. Un-treated (-) and treated (+, 1% O2 treatment for 24hr) HepG2 whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with HIF1 alpha antibody (GTX127309) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX127309_41577_IHC_18121410_579.jpg

GTX127309 IHC-P Image

HIF1 alpha antibody detects HIF1 alpha protein at nucleus on human kidney cancer by immunohistochemical analysis.
Sample: Paraffin-embedded human kidney cancer.
HIF1 alpha antibody (GTX127309) dilution: 1:500.

GTX127309_42662_20170118_IFA_18121410_957.jpg

GTX127309 ICC/IF Image

HIF1 alpha antibody detects HIF1 alpha protein at nucleus by immunofluorescent analysis.
Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: HIF1 alpha protein stained by HIF1 alpha antibody (GTX127309) diluted at 1:500.
Red: Phalloidin, a cytoskeleton marker, diluted at 1:100.
Scale bar = 10 μm.

GTX127309_42123_20151016_IP_18121410_923.jpg

GTX127309 IP Image

Immunoprecipitation of HIF1 alpha protein from HepG2 whole cell extracts treated with 500 μM CoCl2 for 24 hr using 5 μg of HIF1 alpha antibody (GTX127309).
Western blot analysis was performed using HIF1 alpha antibody (GTX127309).
EasyBlot anti-Rabbit IgG (GTX221666-01) was used as a secondary reagent.

GTX127309_42172_20160128_ChIP_18121410_220.jpg

GTX127309 ChIP assay Image

ChIP was performed with HepG2 chromatin extract and 5 μg of either normal rabbit IgG or anti-HIF1 alpha antibody. The precipitated DNA was detected by PCR with primer set targeting to VEGF promoter.

GTX127309_42123_20150924_IFA_2_18121410_338.jpg

GTX127309 ICC/IF Image

HIF1 alpha antibody detects HIF1 alpha protein at nucleus by immunofluorescent analysis.
Sample: NIH/3T3 cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: HIF1 alpha protein stained by HIF1 alpha antibody (GTX127309) diluted at 1:200.
Blue: Hoechst 33342 staining.

GTX127309_42389_20160728_WB_M_CoCl2_18121410_165.jpg

GTX127309 WB Image

Orthogonal

Untreated (–) and treated (+) NIH-3T3 whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with HIF1 alpha antibody (GTX127309) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX127309_41577_WB_CoCl2_18121410_886.jpg

GTX127309 WB Image

Orthogonal

HIF1 alpha antibody detects HIF1 alpha protein by western blot analysis.
A. 30 µg HepG2 whole cell lysate/extract (untreated)
B.30 µg HepG2 whole cell lysate/extract ( 200 μM CoCl2 treatment for 24 hr)
C. 30 µg HepG2 whole cell lysate/extract (500 μM CoCl2 treatment for 24 hr)
7.5% SDS-PAGE
HIF1 alpha antibody (GTX127309) dilution: 1:5000
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX127309_42760_20170316_WB_CoCl2_18121410_159.jpg

GTX127309 WB Image

Orthogonal

Untreated (–) and treated (+) HeLa whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with HIF1 alpha antibody (GTX127309) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX127309_42389_20160526_WB_B_18121410_315.jpg

GTX127309 WB Image

Overexpression

Non-transfected (–) and transfected (+) 293T whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with HIF1 alpha antibody (GTX127309) diluted at 1:5000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

REFERENCE
REVIEW

HIF1 alpha antibody Cat No. GTX127309

Rating ( Average 4.2 based on 15 users reviews)
Application
Western Blot(WB) ( Average 4.4 based on 10 users reviews)
Immunocytochemistry/ Immunofluorescence(ICC/IF) ( Average 4 based on 1 users reviews)
IHC (Paraffin sections)(IHC-P) ( Average 3.7 based on 3 users reviews)
IHC (Frozen sections)(IHC-Fr) ( Average 4 based on 1 users reviews)
Date : Anonymous submitted on 14-Feb-2019
Score :
Application Tested : IHC-Fr
Sample Species : Ms
Sample : Adult cardiac tissue
Blocking : 10% goat serum in PBS-TX100 0.1%, 25°C, 1Hr
Fixation : 4% PFA
Permeabilization /Antigen retrieval : 0.5% Triton-X-100/Citrate Buffer pH6, 0.15Hr
Primary Antibody : 1:50, 4°C, overnightHr
Date : Anonymous submitted on 14-Feb-2019
Score :
Application Tested : IHC-P
Sample Species : Hu
Sample : Renal clear cell carcinoma
Blocking : EnVISION FLEX PEROXIDASE-BLOCKING REAGENT (Dako, Ref. K800)
Fixation : 3.7% formalin
Permeabilization /Antigen retrieval : pH 9 Envision FLEX target retrieval solution, High pH, (Dako Ref. GV800)
Primary Antibody : 1:100, 25°C, 1Hr
Date : Anonymous submitted on 30-Nov-2017
Score :
Application Tested : WB
Sample Species : Hu
Sample : HeLa cells
Amount used : 10 μg
Blocking : ATTO EzBlock Chemi, 25°C, 0.5Hr
Primary Antibody : 1:5000, 4°C, overnightHr
Date : Anonymous submitted on 22-Nov-2017
Score :
Application Tested : IHC-P
Sample : HeLa xenograft
Blocking : 10% goat serum, 25°C, 0.08Hr
Fixation : Formalin Neutral Buffer
Permeabilization /Antigen retrieval : 0.2% Brij35 in PBS/Dako S2369
Primary Antibody : 1:100, 25°C, 0.6Hr
Date : Anonymous submitted on 05-Oct-2017
Score :
Application Tested : WB
Sample Species : Hu
Sample : OVSAYO cells
Blocking : 3% skim milk, 25°C, 0.5Hr
Primary Antibody : 1:2000, 4°C, 16Hr
Lane Description : Lane 1: Vehicle, Lane 2: 500 μM Cocl2 treatment for 4 hr
Date : Anonymous submitted on 05-Oct-2017
Score :
Application Tested : WB
Sample Species : Hu
Sample : OVSAYO cells
Blocking : 3% skim milk, 25°C, 0.5Hr
Primary Antibody : 1:2000, 4°C, 16Hr
Lane Description : Lane 1: Normoxia, Lane 2: 1% O2 treatment for 4 hr
Date : Anonymous submitted on 22-Sep-2016
Score :
Application Tested : WB
Sample Species : Hu
Sample : dental pulp stem cells
Amount used : 1x10^4 cells
Blocking : PVDF blocking reagent, 25°C, 0.25Hr
Primary Antibody : 1:1000, 4°C, overnightHr
Date : Anonymous submitted on 02-Mar-2016
Score :
Application Tested : WB
Sample Species : Hu
Sample : THP-1 cells
Amount used : 20μg
Blocking : 5% milk in TBST, 25°C, 1Hr
Primary Antibody : 1:2000, 4°C, overnightHr
Lane Description : Lane 1:THP-1 cells (250ug/ml TPA treatment for 4h), Lane 2:THP-1 cells (250ug/ml TPA and 1 ug/ml LPS treatment for 4h)
Date : Anonymous submitted on 29-Jan-2016
Score :
Application Tested : ICC/IF
Sample Species : Hu
Sample : A549 cells
Fixation : 4% PFA, 2Hr
Permeabilization /Antigen retrieval : Citrate buffer, 95°C, 20minHr
Primary Antibody : 1:1000, 25°C, 1Hr
Figure Note : A: A549 cells, B: A549 cells (1mM DMOG treatment for 3.5 h)
Date : Anonymous submitted on 29-Jan-2016
Score :
Application Tested : IHC-P
Sample Species : Hu
Sample : brain tissue
Fixation : 4% PFA, 2Hr
Permeabilization /Antigen retrieval : Citrate buffer, 95°C, 20minHr
Primary Antibody : 1:1000, 25°C, 1Hr
Figure Note : A:Human glioblastoma tissue, B:Human healthy brain tissue
Date : Anonymous submitted on 30-Dec-2015
Score :
Application Tested : WB
Sample Species : Hu
Sample : meniscal fibrochondrocytes
Amount used : 15μg
Blocking : BlockingPRO Blocking buffer, 25°C, 1Hr
Primary Antibody : 1:1000, 4°C, overnightHr
Lane Description : Lane 1: meniscal fibrochondrocytes (normoxia), Lane 2: meniscal fibrochondrocytes (1% O2 treatment for 6hr)
Date : Anonymous submitted on 08-Dec-2014
Score :
Application Tested : WB
Sample Species : Hu
Sample : endometriotic stromal cells
Amount used : 20μg
Blocking : 5% non-fat milk, 25°C, 1Hr
Primary Antibody : 1:5000, 25°C, 1Hr
Lane Description : Lane A:DMSO treatment (24hr) ; Lane B:0.1uM DFO treatment (24hr) ; Lane C:1uM DFO treatment (24hr)
Date : Anonymous submitted on 14-Aug-2014
Score :
Application Tested : WB
Sample Species : Hu
Sample : HeLa cells
Amount used : 20μg
Blocking : 25°C, 1Hr
Primary Antibody : 1:1000, 4°C, 12Hr
Lane Description : Lane A: HeLa cell lysate(normoxia), Lane B: HeLa cell lysate (500μM CoCl2 treatment)
Date : Anonymous submitted on 27-Aug-2013
Score :
Application Tested : WB
Sample Species : Hu
Sample : A431 cells
Amount used : 20μg
Blocking : 5% milk in TBST, 37°C, 1Hr
Primary Antibody : 1:1000, 37°C, 1Hr
Lane Description : A: A431 whole cell lysate under normoxia, B: A431 whole cell lysate under hypoxia
Date : Anonymous submitted on 16-Oct-2012
Score :
Application Tested : WB
Sample Species : Hu
Sample : A549 cells
Amount used : 40μg
Blocking : 3% milk , 25°C, 1Hr
Primary Antibody : 1:10000, 4°C, overnightHr
Application Reference
SDS
PBS.pdf
Glycerol.pdf
Proclin.pdf