APPLICATION
Application Note
*Optimal dilutions/concentrations should be determined by the researcher.
| Application |
Dilution |
| 1:500-1:3000 |
| 1:100-1:1000 |
| 1:100-1:1000 |
| Assay dependent |
| 1:100-1:570 |
| Assay dependent |
Not tested in other applications.
Calculated MW
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
0.26 mg/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
Hypoxia Inducible Factor 1 Subunit Alpha,Hif-1-Alpha,Hif-1A,Hif-1Alpha,Hif1,Hif1-Alpha,Mop1,Pasd8,Bhlhe78,Hif1A
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
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GTX127309 WB Image
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.
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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
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GTX127309 WB Image
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.
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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.
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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.
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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.
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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.
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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.
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GTX127309 WB Image
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.
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GTX127309 WB Image
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.
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GTX127309 WB Image
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.
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GTX127309 WB Image
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.
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REFERENCE
REVIEW
HIF1 alpha antibody
Cat No. GTX127309
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Rating
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( Average 4.2 based on 15 users reviews)
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Application
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Western Blot(WB)
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( Average 4.4 based on 10 users reviews)
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Immunocytochemistry/ Immunofluorescence(ICC/IF)
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( Average 4 based on 1 users reviews)
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IHC (Paraffin sections)(IHC-P)
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( Average 3.7 based on 3 users reviews)
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IHC (Frozen sections)(IHC-Fr)
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( Average 4 based on 1 users reviews)
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Date :
Anonymous submitted on 14-Feb-2019
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
