HIF-1 alpha Recombinant Rabbit Monoclonal Antibody [PSH07-00]
cat.: HA722778
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | PSH07-00 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% 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: | Synthetic peptide within human HIF-1 alpha aa 441-490. |
| Positive control: | HeLa cell lysate, HeLa treated with 0.5mM CoCl2 for 6 hours cell lysate, HepG2 cell lysate, HepG2 treated with 100μM CoCl2 for 4 hours cell lysate, C2C12 cell lysate, C2C12 treated with 100μM CoCl2 for 4 hours cell lysate, HeLa cells treated with 0.5mM CoCl2 for 6 hours, C2C12 cells treated with 200μM CoCl2 for 4 hours. |
| Subcellular location: | Cytoplasm, Nucleus, Nucleus speckle. |
| Recommended Dilutions:
WB IF-Cell IHC-P |
1:5,000 1:100 1:1,000 |
| 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 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
|
Fig1:
Western blot analysis of HIF-1 alpha on different lysates with Rabbit anti-HIF-1 alpha antibody (HA722778) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 0.5mM CoCl2 for 6 hours cell lysate Lane 3: HepG2 cell lysate Lane 4: HepG2 treated with 100μM CoCl2 for 4 hours cell lysate Lane 5: C2C12 cell lysate Lane 6: C2C12 treated with 100μM CoCl2 for 4 hours cell lysate Lysates/proteins at 30 µg/Lane. Predicted band size: 93 kDa Observed band size: 120 kDa Exposure time: 20 seconds; ECL: K1802; 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 (HA722778) at 1/5,000 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. |
|
Fig2:
Immunocytochemistry analysis of HeLa cells treated with 0.5mM CoCl2 for 6 hours labeling HIF-1 alpha with Rabbit anti-HIF-1 alpha antibody (HA722778) 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 (HA722778) 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. |
|
Fig3:
Immunocytochemistry analysis of C2C12 cells treated with 200μM CoCl2 for 4 hours labeling HIF-1 alpha with Rabbit anti-HIF-1 alpha antibody (HA722778) 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 (HA722778) 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. |
|
Fig4:
Immunohistochemical analysis of paraffin-embedded HeLa cells treated with 0.5mM CoCl2 for 6 hours with Rabbit anti-HIF-1 alpha antibody (HA722778) 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 (HA722778) 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.