HIF-1 Alpha Rabbit Polyclonal Antibody

• Catalog Number: orb500743
• Category: Antibodies
• Description: HIF-1 Alpha Rabbit Polyclonal Antibody
• Target: HIF1A
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human
• Predicted Reactivity: Gallus, Mouse, Rat
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: Preservative: 0.02% Proclin300, Constituents: 1% rAlbumin, 0.01M PBS, pH 7.4.
• Immunogen: KLH conjugated synthetic peptide derived from human HIF-1 Alpha (661-760/826aa)
• UniProt ID: Q16665
• MW: 92 kDa
• Tested applications: FC, WB
• Dilution range: WB=1:500-2000, Flow-Cyt=1μg/Test
• Antibody Type: Primary Antibody
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: ARNT interacting protein; ARNT-interacting protein
• Note: For research use only

Sample: Lane 1: Hela (Human) Cell Lysate at 30 ug, Lane 2: A431 (Human) Cell Lysate at 30 ug, Lane 3: U251 (Human) Cell Lysate at 30 ug, Lane 4: HepG2 (Human) Cell Lysate at 30 ug, Primary: Anti-HIF-1 Alpha (orb500743) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 92 kD, Observed band size: 120 kD.

Sample: Lane 1: Recombinant human HIF-1 Alpha protein, N-Trx-His, Primary: Anti-HIF-1 Alpha (orb500743) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 92 kDa, Observed band size: 52 kDa.

Blank control (blue line): Hela (fixed with 80% methanol (5 min at -20°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature). Primary Antibody (green line): Rabbit Anti-HIF-1 Alpha antibody (orb500743), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, dilution: 1 µg/Test.

Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (orb500743) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human kidney), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (orb500743) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat kidney), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (orb500743) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat liver), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (orb500743) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Tissue/Cell: U-87MG cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (HIF-1 Alpha) polyclonal Antibody, Unconjugated (orb500743) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.