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HIF-1 alpha Recombinant Rabbit Monoclonal Antibody [PSH07-00]

Usd: 385

Specification

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Catalog# HA722778

HIF-1 alpha Recombinant Rabbit Monoclonal Antibody [PSH07-00]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

HIF-1 alpha Recombinant Rabbit Monoclonal Antibody [PSH07-00]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human HIF-1 alpha aa 441-490.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IHC-P

Target Molecular Weight

Predicted band size: 93 kDa

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.

Conjugation

unconjugated

Clone Number

PSH07-00

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000

Target

Function

Hypoxia-inducible factor 1-alpha, also known as HIF-1-alpha, is a subunit of a heterodimeric transcription factor hypoxia-inducible factor 1 (HIF-1) that is encoded by the HIF1A gene. The Nobel Prize in Physiology or Medicine 2019 was awarded for the discovery of HIF. HIF1A is a basic helix-loop-helix PAS domain containing protein, and is considered as the master transcriptional regulator of cellular and developmental response to hypoxia. The dysregulation and overexpression of HIF1A by either hypoxia or genetic alternations have been heavily implicated in cancer biology, as well as a number of other pathophysiologies, specifically in areas of vascularization and angiogenesis, energy metabolism, cell survival, and tumor invasion. Two other alternative transcripts encoding different isoforms have been identified.

Background References

1. Korbecki J et al. Chronic and Cycling Hypoxia: Drivers of Cancer Chronic Inflammation through HIF-1 and NF-κB Activation: A Review of the Molecular Mechanisms. Int J Mol Sci. 2021 Oct

2. Infantino V et al. Cancer Cell Metabolism in Hypoxia: Role of HIF-1 as Key Regulator and Therapeutic Target. Int J Mol Sci. 2021 May

Subcellular Location

Cytoplasm, Nucleus, Nucleus speckle.

UNIPROT

SwissProt: Q16665 Human

SwissProt: Q61221 Mouse

Synonyms

ARNT interacting protein antibody

ARNT-interacting protein antibody

Basic helix loop helix PAS protein MOP1 antibody

Basic-helix-loop-helix-PAS protein MOP1 antibody

bHLHe78 antibody

Class E basic helix-loop-helix protein 78 antibody

HIF 1A antibody

HIF 1alpha antibody

HIF-1-alpha antibody

HIF-1alpha antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of HIF-1 alpha on different lysates with Rabbit anti-HIF-1 alpha antibody (<a href="/products/HA722778" style="font-weight: bold;text-decoration: underline;">HA722778</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 0.5mM CoCl2 for 6 hours cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: HepG2 treated with 100μM CoCl2 for 4 hours cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: C2C12 treated with 100μM CoCl2 for 4 hours cell lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 93 kDa<br />Observed band size: 120 kDa<br /><br />Exposure time: 20 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722778" style="font-weight: bold;text-decoration: underline;">HA722778</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with 0.5mM CoCl2 for 6 hours labeling HIF-1 alpha with Rabbit anti-HIF-1 alpha antibody (<a href="/products/HA722778" style="font-weight: bold;text-decoration: underline;">HA722778</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/HA722778" style="font-weight: bold;text-decoration: underline;">HA722778</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of C2C12 cells treated with 200μM CoCl2 for 4 hours labeling HIF-1 alpha with Rabbit anti-HIF-1 alpha antibody (<a href="/products/HA722778" style="font-weight: bold;text-decoration: underline;">HA722778</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/HA722778" style="font-weight: bold;text-decoration: underline;">HA722778</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunohistochemical analysis of paraffin-embedded HeLa cells treated with 0.5mM CoCl2 for 6 hours with Rabbit anti-HIF-1 alpha antibody (<a href="/products/HA722778" style="font-weight: bold;text-decoration: underline;">HA722778</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722778" style="font-weight: bold;text-decoration: underline;">HA722778</a>) 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.

    ☑ Cell treatment (CT)

    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) (high pressure) 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.

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