HIF-1 Alpha Polyclonal Antibody

$Tissue\/cell: U-87MG cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (HIF-1 Alpha) polyclonal Antibody, Unconjugated (bs-20399R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei$

Tissue\/cell: U-87MG cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (HIF-1 Alpha) polyclonal Antibody, Unconjugated (bs-20399R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei

$Lane 1: Human A431 cell lysates; Lane 2: Human U-2OS cell lysates; Lane 3: Human HepG2 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-20399R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Human A431 cell lysates; Lane 2: Human U-2OS cell lysates; Lane 3: Human HepG2 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-20399R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

$Lane 1: Human Hela cell lysates; Lane 2: Human A431 cell lysates; Lane 3: Human U251 cell lysates; Lane 4: Human HepG2 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-20399R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Human Hela cell lysates; Lane 2: Human A431 cell lysates; Lane 3: Human U251 cell lysates; Lane 4: Human HepG2 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-20399R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

$Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-20399R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-20399R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-20399R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-20399R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Paraformaldehyde-fixed, paraffin embedded (human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-20399R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-20399R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-20399R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-20399R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$Lane 1: Recombinant human TNFSF13B protein, N-His(bs-43649P) probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-20399R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.$

Lane 1: Recombinant human TNFSF13B protein, N-His(bs-43649P) probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-20399R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.

$The HepG2 (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-HIF-1 Alpha antibody (bs-20399R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-40295G-FITC): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

The HepG2 (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-HIF-1 Alpha antibody (bs-20399R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-40295G-FITC): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)

Reactivity

Human
Rat

Predicted Reactivity

Mouse
Chicken

Overview
Catalog #	bs-20399R
Product Name	HIF-1 Alpha Polyclonal Antibody
Applications	WB ELISA FCM IHC-P IHC-F IF(IHC-P) IF(IHC-F) IF(ICC)
Reactivity	Human, Rat
Predicted Reactivity	Mouse, Chicken
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic peptide derived from human HIF-1 Alpha
Immunogen Range	661-760/826
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	3091
Swiss Prot	Q16665
Subcellular location	Cytoplasm, Nucleus
Synonyms	HIF-1-alpha; hypoxia-inducible factor 1, alpha subunit isoform 1; Hypoxia-inducible factor 1 alpha; HIF-1 alpha; HIF1 alpha; HIF-1a; HIF-1alpha; ARNT-interacting protein; Member of PAS protein1; Basic-helix-loop-helix-PAS protein MOP1; Hypoxia-inducible factor -1 Alpha.
Background	Functions as a master transcriptional regulator of theadaptive response to hypoxia. Under hypoxic conditions activatesthe transcription of over 40 genes, including, erythropoietin,glucose transporters, glycolytic enzymes, vascular endothelialgrowth factor, and other genes whose protein products increaseoxygen delivery or facilitate metabolic adaptation to hypoxia.Plays an essential role in embryonic vascularization, tumorangiogenesis and pathophysiology of ischemic disease. Binds to coreDNA sequence 5'-[AG]CGTG-3' within the hypoxia response element(HRE) of target gene promoters. Activation requires recruitment oftranscriptional coactivators such as CREBPB and EP300. Activity isenhanced by interaction with both, NCOA1 or NCOA2. Interaction withredox regulatory protein APEX seems to activate CTAD andpotentiates activation by NCOA1 and CREBBP.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200