HIF1 alpha Rabbit Polyclonal Antibody
cat.: ER1802-41
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Rat, Mouse, Rabbit, Pig, Cow, Dog
Applications: WB, IHC-P, FC, ICC, IF, ELISA
Clonality: Polyclonal
Form: Liquid
Storage condition: Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 92 kDa
Isotype: IgG
Immunogen: Synthetic peptide within C-terminal human HIF1 alpha.
Subcellular location: Cytoplasm. Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  FC
  ICC
  IF
  ELISA
1:500-1:2,000
1:100-1:500
1μg/Test
1:100
1:100-1:500
1:5,000-1:10,000
Uniprot #: SwissProt: Q16665 Human | Q61221 Mouse
Alternative names: ARNT interacting protein ARNT-interacting protein Basic helix loop helix PAS protein MOP1 Basic-helix-loop-helix-PAS protein MOP1 bHLHe78 Class E basic helix-loop-helix protein 78 HIF 1A HIF 1alpha HIF-1-alpha HIF1 A HIF1 Alpha HIF1 HIF1-alpha HIF1A HIF1A_HUMAN Hypoxia inducible factor 1 alpha Hypoxia inducible factor 1 alpha isoform I.3 Hypoxia inducible factor 1 alpha subunit Hypoxia inducible factor 1 alpha subunit basic helix loop helix transcription factor Hypoxia inducible factor 1, alpha subunit (basic helix loop helix transcription factor) Hypoxia inducible factor1alpha Hypoxia-inducible factor 1-alpha Member of PAS protein 1 Member of PAS superfamily 1 Member of the PAS Superfamily 1 MOP 1 MOP1 PAS domain-containing protein 8 PASD 8 PASD8
Images
ER1802-41_1.jpg Fig1: Blank control (blue line): Hela (blue).
Primary Antibody (green line): Rabbit Anti-HIF1 alpha antibody (ER1802-41)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 80% methanol (5 min at -20℃) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
ER1802-41_2.jpg Fig2: Tissue/cell: rat lung tissue(Smoking); 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-HIF1 alpha Polyclonal Antibody, Unconjugated(ER1802-41) 1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1802-41_3.jpg Fig3: Tissue/cell: human cervical carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-HIF1 alpha Polyclonal Antibody, Unconjugated(ER1802-41) 1:300, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1802-41_4.jpg Fig4: 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-HIF1 alpha (ER1802-41) at 1/1,000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20,000 dilution
Predicted band size: 92 kD
Observed band size: 120 kD
ER1802-41_5.jpg Fig5: Blank control:Mouse spleen.
Primary Antibody (green line): Rabbit Anti-HIF1 alpha antibody (ER1802-41)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
ER1802-41_6.jpg Fig6: Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37℃ for 20 min; Antibody incubation with (HIF1 alpha) polyclonal Antibody, Unconjugated (ER1802-41) 1:100, 90 minutes at 37℃; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37℃ for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
ER1802-41_7.jpg Fig7: Sample:
Lane 1: U87MG (Human) Cell Lysate at 30 ug
Lane 2: Hela (Human) Cell Lysate at 30 ug
Lane 3: A431 (Human) Cell Lysate at 30 ug
Lane 4: U251 (Human) Cell Lysate at 30 ug
Primary: Anti-HIF1 alpha (ER1802-41) at 1/1,000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20,000 dilution
Predicted band size: 92 kD
Observed band size: 120 kD
ER1802-41_8.jpg Fig8: Paraformaldehyde-fixed, paraffin embedded (rat stomach); 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℃ for 30min; Antibody incubation with (HIF1 alpha) Polyclonal Antibody, Unconjugated (ER1802-41) at 1:200 overnight at 4℃, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1802-41_9.jpg Fig9: 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℃ for 30min; Antibody incubation with (HIF1 alpha) Polyclonal Antibody, Unconjugated (ER1802-41) at 1:200 overnight at 4℃, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1802-41_10.jpg Fig10: Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37℃ for 20 min; Antibody incubation with (HIF1 alpha) polyclonal Antibody, Unconjugated (ER1802-41) 1:100, 90 minutes at 37℃; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37℃ for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.