HIF-1 Alpha Polyclonal Antibody

Formalin-fixed and paraffin embedded human cervical carcinoma labeled Anti-HIF-1-Alpha Polyclonal Antibody (bs-0737R), Unconjugated at 1:300, followed by conjugation to the secondary antibody and DAB staining

L1 mouse heart lysates, L2 mouse large intestine lysates probed with Anti HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in 37˚C. Predicted band and observed band size: 92kD

KLN 205 cells labeled with Rabbit HIF-1 Alpha Polyclonal Antibody Polyclonal Antibody, Unconjugated (bs-0737R) followed by a FITC Conjugated Secondary Antibody.

Image kindly submitted by Edita Aksamitiene as part of the Free Sample Program. Rat brain and spleen lysates probed with Rabbit Anti-HIF-1 Alpha Polyclonal Antibody (bs-0737R) at 1:1000.

HL-60 cell lysates probed with Anti-HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:300 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at 37˚C.

HeLa cells probed with HIF-1 Alpha Polyclonal Antibody, unconjugated (bs-0737R) at 1:100 dilution for 30 minutes compared to control cells (blue) and isotype control (orange)

Lane 1: Hela cell lysates; Lane 2: A431 cell lysates; Lane 3: U251 cell lysates; Lane 4: HepG2 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Hela cells(black) were fixed with 70% ice-cold methanol overnight at 4\u2103,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with \r\nHIF-1 Alpha Polyclonal Antibody(bs-0737R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Hela cells(black) were fixed with 70% ice-cold methanol overnight at 4\u2103,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with \r\nHIF-1 Alpha Polyclonal Antibody(bs-0737R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (HIF-1 Alpha) polyclonal Antibody, Unconjugated (bs-0737R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.$

Hela 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-0737R) 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 U-87MG cell lysates; Lane 2: Human Hela cell lysates; Lane 3: Human A431 cell lysates; Lane 4: Human U251 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Mouse spleen cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HIF-1 Alpha Antibody(bs-0737R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Mouse spleen cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HIF-1 Alpha Antibody(bs-0737R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

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-0737R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Applications

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

Reactivity

Human
Mouse
Rat
Rabbit
Others

Predicted Reactivity

Overview
Catalog #	bs-0737R
Product Name	HIF-1 Alpha Polyclonal Antibody
Applications	WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC), IP
Reactivity	Human, Mouse, Rat, Rabbit, Others
Predicted Reactivity	Dog, Cow, Pig
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic peptide derived from human of human HIF-1 Alpha
Immunogen Range	341-450/826
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 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	3091
Swiss Prot	Q16665
Subcellular location	Cytoplasm, Nucleus
Synonyms	HIF1; MOP1; PASD8; HIF-1A; bHLHe78; HIF-1alpha; HIF1-ALPHA; Hypoxia-inducible factor 1-alpha; HIF-1-alpha; ARNT-interacting protein; Basic-helix-loop-helix-PAS protein MOP1; Class E basic helix-loop-helix protein 78; Member of PAS protein 1; PAS domain-containing protein 8; HIF1A
Background	Functions as a master transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions, activates the transcription of over 4 genes, including erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, HILPDA, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Activation requires recruitment of transcriptional coactivators such as CREBPB and EP3. Activity is enhanced by interaction with both, NCOA1 or NCOA2. Interaction with redox regulatory protein APEX seems to activate CTAD and potentiates activation by NCOA1 and CREBBP. Involved in the axonal distribution and transport of mitochondria in neurons during hypoxia.
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
IP	1-2ug