HIF-2 alpha Recombinant Rabbit Monoclonal Antibody [JB24-42]
cat.: ET7107-32
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, FC
Clonality: Monoclonal
Clone number: JB24-42
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 96 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human HIF-2 alpha aa 61-110 / 870.
Positive control: Mouse lung tissue lysate, Siha cell lysate, rat lung tissue, human breast tissue, human placenta tissue, mouse colon tissue, HUVEC.
Subcellular location: Nucleus, Nucleus speckle.
Recommended Dilutions:
  WB
  IHC-P
  FC
1:500-1:2,000
1:50-1:1,000
1:50-1:100
Uniprot #: SwissProt: Q99814 Human | P97481 Mouse | Q9JHS1 Rat
Alternative names: Basic helix loop helix PAS protein MOP2 Basic-helix-loop-helix-PAS protein MOP2 bHLHe73 Class E basic helix-loop-helix protein 73 ECYT4 Endothelial PAS domain containing protein 1 Endothelial pas domain protein 1 Endothelial PAS domain-containing protein 1 EPAS 1 EPAS-1 EPAS1 EPAS1_HUMAN HIF 1 alpha like factor HIF 2 alpha HIF-1-alpha-like factor HIF-2-alpha HIF2-alpha HIF2A HLF Hypoxia inducible factor 2 alpha Hypoxia inducible factor 2 alpha subunit Hypoxia-inducible factor 2-alpha Member of PAS protein 2 Member of pas superfamily 2 MOP 2 MOP2 PAS domain-containing protein 2 PASD2
Images
ET7107-32_1.jpg Fig1: Western blot analysis of HIF-2 alpha on different lysates with Rabbit anti-HIF-2 alpha antibody (ET7107-32) at 1/500 dilution.

Lane 1: Mouse lung tissue lysate (20 µg/Lane)
Lane 2: Siha cell lysate (10 µg/Lane)

Predicted band size: 96 kDa
Observed band size: 110 kDa

Exposure time: 1 minute;

8% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-32) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET7107-32_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded rat lung tissue using anti-HIF-2 alpha antibody. Counter stained with hematoxylin.
ET7107-32_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-HIF-2 alpha antibody (ET7107-32) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-32) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET7107-32_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-HIF-2 alpha antibody. Counter stained with hematoxylin.
ET7107-32_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-HIF-2 alpha antibody. Counter stained with hematoxylin.
ET7107-32_6.jpg Fig6: Flow cytometric analysis of HUVEC cells with HIF-2 antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
ET7107-32_7.jpg Fig7: Immunocytochemistry analysis of Siha cells labeling HIF-2 alpha with Rabbit anti-HIF-2 alpha antibody (ET7107-32) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HIF-2 alpha antibody (ET7107-32) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.