HIF-1 alpha Recombinant Rabbit Monoclonal Antibody [JE75-33]
cat.: HA721997
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: JE75-33
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. 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: 93 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human HIF-1 alpha aa 251-550 / 826.
Positive control: HeLa treated with 250μM CoCl2 for 6 hours cell lysate, HepG2 treated with 250μM CoCl2 for 6 hours cell lysate, HeLa cells treated with or without 500μM CoCl2 for 24 hours, human kidney tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location: Cytoplasm, Nucleus, Nucleus speckle.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P

1:1,000
1:100
1:200-1:500
Uniprot #: SwissProt: Q16665 Human | Q61221 Mouse | O35800 Rat
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 HIF-1alpha HIF-alpha HIF1 A HIF1 Alpha HIF1 HIF1-alpha HIF1A HIF1A_HUMAN hifla 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
HA721997_1.jpg Fig1: Western blot analysis of HIF-1 alpha on different lysates with Rabbit anti-HIF-1 alpha antibody (HA721997) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 250μM CoCl2 for 6 hours cell lysate
Lane 3: HepG2 cell lysate
Lane 4: HepG2 treated with 250μM CoCl2 for 6 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 93 kDa
Observed band size: 120 kDa

Exposure time: 1minute 2 seconds;

4-20% 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 (HA721997) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA721997_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells treated with or without 500μM CoCl2 for 24 hours labeling HIF-1 alpha with Rabbit anti-HIF-1 alpha antibody (HA721997) 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-1 alpha antibody (HA721997) 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.
HA721997_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-HIF-1 alpha antibody (HA721997) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721997) at 1/200 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.
HA721997_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-HIF-1 alpha antibody (HA721997) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721997) at 1/200 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.
HA721997_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-HIF-1 alpha antibody (HA721997) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721997) at 1/500 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.