HIF1AN Recombinant Antibody

$4% Paraformaldehyde-fixed Jurkat (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (HIF1AN) monoclonal Antibody, unconjugated (bsm-61905R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.$

4% Paraformaldehyde-fixed Jurkat (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (HIF1AN) monoclonal Antibody, unconjugated (bsm-61905R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

$Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Prostate; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Prostate; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with HIF1AN Monoclonal Antibody, Unconjugated (bsm-61905R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$The Jurkat (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-HIF1AN antibody (bsm-61905R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

The Jurkat (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.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-HIF1AN antibody (bsm-61905R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

$25 ug total protein per lane of various lysates (see on figure) probed with HIF1AN monoclonal antibody, unconjugated (bsm-61905R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with HIF1AN monoclonal antibody, unconjugated (bsm-61905R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Applications

WB
IHC-P
IHC-F
IF
Flow-Cyt
ICC/IF

Reactivity

Human

Overview
Catalog #	bsm-61905r-100ul
Product Name	HIF1AN Recombinant Antibody
Applications	WB, IHC-P, IHC-F, IF, Flow-Cyt, ICC/IF
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic peptide derived from human HIF 1 alpha inhibitor
Clonality	Recombinant
Isotype	IgG
Concentration	1mg/ml
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	55662
Swiss Prot	Q9NWT6
Subcellular location	Nucleus
Synonyms	FIH 1; FIH1; HIF1AN.
Background	Hydroxylates HIF-1 alpha at 'Asn-803' in the C-terminal transactivation domain (CAD). Functions as an oxygen sensor and, under normoxic conditions, the hydroxylation prevents interaction of HIF-1 with transcriptional coactivators including Cbp/p300-interacting transactivator. Involved in transcriptional repression through interaction with HIF1A, VHL and histone deacetylases. Hydroxylates specific Asn residues within ankyrin repeat domains (ARD) of NFKB1, NFKBIA, NOTCH1, ASB4, PPP1R12A and several other ARD-containing proteins.
Application Dilution
WB	1:500-2000
IHC-P	1:200-500
IHC-F	1:200-500
IF	1:200-500
Flow-Cyt	1ug/test
ICC/IF	1:100-200