HIF 2 alpha Rabbit Polyclonal Antibody

• Catalog Number: orb10804
• Category: Antibodies
• Description: HIF 2 alpha Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, WB
• Predicted Reactivity: Human
• Reactivity: Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from mouse HIF 2 Alpha (1-130/874aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, ICC/IF=1:50-200, Flow-Cyt=1:20-100
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 96 kDa
• Target: EPAS1
• UniProt ID: P97481
• RRID: AB_10928187
• 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.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Basic helix loop helix PAS protein MOP2; Basic-hel
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue line): Hep G2 (blue). Primary Antibody (green line): Rabbit Anti-HIF 2 alpha antibody (orb10804), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, Dilution: 1 µg/Test. Protocol, The cells were fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% methanol for 20 min at -20°C. 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 20000 events was performed.

Blank control: RSC96 (blue). Primary Antibody: Rabbit Anti-HIF 2α antibody (orb10804), Dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions, Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA. Protocol, The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Antibody (orb10804, 51g /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of orb10804 at 1/200 dilution for 30 min on ice. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain), 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 2 alpha) Polyclonal Antibody, Unconjugated (orb10804) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: Lung (Rat) lysate at 30 ug, Primary: Rabbit Anti-HIF 2α (orb10804) at 1:200 dilution, Secondary: HRP conjugated Goat Anti-Rabbit IgG (orb572747) at 1:3000 dilution, Predicted band size: 96kD, Observed band size: 96kD.

Tissue/Cell: human glioma tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-HIF 2α Polyclonal Antibody, Unconjugated (orb10804) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/Cell: mouse lymph node, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-HIF 2αPolyclonal Antibody, Unconjugated (orb10804) 1:200, overnight at 4°C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (orb868589) used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 ug/ml, blue) was used to stain the cell nuclei.