A Short Overview of Hypoxia Markers

Hypoxia markers are cellular molecules which are expressed at greater levels when the cell or tissue is experiencing hypoxic conditions.

Some of them are listed below.

HIF-1α

When the cell is hypoxic, HIF-1a becomes a stable molecule and migrates into the nucleus. Here, it acts as a transcription regulator of certain genes which express proteins that increase the amount of oxygen delivered from the blood to the peripheral tissues or help the cells adapt faster and better in their metabolic processes to the presence of low oxygen levels.

HIF-1a staining of HeLa cells. Anti-HIF-1a antibody [EPR16897] (ab179483) was used at 1/50 dilution with goat anti-rabbit IgG (Alexa Fluor® 488) secondary antibody. Cells were treated with 0.6 nM CoCl2 for 6 hours.

Figure 1. HIF-1a staining of HeLa cells. Anti-HIF-1a antibody [EPR16897] (ab179483) was used at 1/50 dilution with goat anti-rabbit IgG (Alexa Fluor® 488) secondary antibody. Cells were treated with 0.6 nM CoCl2 for 6 hours.

Application Recommended product
Western blot, immunocytochemistry Anti-HIF-1a antibody (ab179483)
ELISA Human SimpleStep ELISA® kit (ab171577)
Positive control Recombinant human HIF-1a protein (ab154478)
Activity assay HIF-1a transcription factor assay kit (ab133104)

BNIP3

HIF-1a can induce the expression of BNIP3 when the cell is hypoxic, and it is linked to various phenomena, such as autophagy mediated by hypoxia, cell survival mechanisms and progression of tumors1.

BNIP3 staining of laryngeal squamous cell carcinoma tissue. Anti-BNIP3 mouse monoclonal antibody (ab10433) was used at 1/100 dilution and visualized by DAB.

Figure 2. BNIP3 staining of laryngeal squamous cell carcinoma tissue. Anti-BNIP3 mouse monoclonal antibody (ab10433) was used at 1/100 dilution and visualized by DAB.

Application Recommended product
Various Anti-BNIP antibody [ANa40] (ab10433)
ELISA Human HIF-1a + BNIP3 hypoxia in cell ELISA kit (ab129733)
Flow cytometry HIF-1a + BNIP3 hypoxia response human flow cytometry kit (ab126585)

PDK1

PDK1 (pyruvate dehydrogenase kinase 1) is a molecule whose expression is dependent upon HIF-1a expression. It is therefore, a marker of hypoxia, and it is responsible for moving glucose through the glycolysis pathway - an anerobic metabolic route - so that ATP can still be produced for the body’s energy needs2.

PDK1 staining of NIH/3T3 cells. Anti-mitochondrial pyruvate dehydrogenase kinase 1 antibody (ab202468) was used at a dilution of 1/1000 with goat anti-rabbit IgG (Alexa Fluor 488) secondary antibody (ab150077). Cells were counterstained with DAPI (blue) and  tubulin also stained (red).

Figure 3. PDK1 staining of NIH/3T3 cells. Anti-mitochondrial pyruvate dehydrogenase kinase 1 antibody (ab202468) was used at a dilution of 1/1000 with goat anti-rabbit IgG (Alexa Fluor 488) secondary antibody (ab150077). Cells were counterstained with DAPI (blue) and  tubulin also stained (red).

Application Recommended product
Western blot, ICC, IP, flow cytometry Anti-mitochondrial pyruvate dehydrogenase kinase 1 antibody (ab202468)
ELISA Human HIF-1a + PDK1 hypoxia in cell ELISA kit (ab125299)
Flow cytometry HIF-1a + PDK1 hypoxia response human flow cytometry kit (ab126700)
Positive control Recombinant human mitochondrial pyruvate dehydrogenase kinase 1 protein (ab125580)

GLUT1

The GLUT1 protein is a glucose transporter that is downstream of HIF-1a, and is therefore expressed more actively in hypoxia3.

GLUT1 staining of HEPg2 cells. Anti-glucose transporter GLUT antibody (ab115730) was used at a dilution of 1/100 with Alexa Fluor® 488 goat anti-rabbit secondary antibody (ab150077). Cells were counterstained with DAPI and tubulin was also stained (red).

Figure 4. GLUT1 staining of HEPg2 cells. Anti-glucose transporter GLUT antibody (ab115730) was used at a dilution of 1/100 with Alexa Fluor® 488 goat anti-rabbit secondary antibody (ab150077). Cells were counterstained with DAPI and tubulin was also stained (red).

Application Recommended product
Western blot, ICC/IHC, flow cytometry Anti-glucose transporter GLUT antibody (ab115730)
ELISA Human HIF-1a + GLUT hypoxia in cell ELISA kit (ab125298)
Flow cytometry HIF-1a + GLUT1 hypoxia response human flow cytometry kit (ab126584)

References

  1. Bellot G, Garcia-Medina, Gounon P, Chiche J, Roux D, Pouysségur J, Mazure NM. Hypoxia-induced autophagy is mediated through hypoxia-inducible factor induction of BNIP3 and BNIP3L via their BH3 domains. Mol Cell Biol. 10, 2570–81 (2009).
  2. Kim JW, Tchernyshyov I, Semenza GL, Dang CV. HIF-1-mediated expression of pyruvate dehydrogenase kinase: a metabolic switch required for cellular adaptation to hypoxia. Cell Metab. 3, 177–85 (2006).
  3. Chen C, Pore N, Behrooz A, Ismail-Beigi F, Maity A. Regulation of glut1 mRNA by hypoxia-inducible factor-1. Interaction between H-ras and hypoxia. J Biol Chem. 276, 9519–25 (2000).

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Last updated: Apr 1, 2019 at 6:15 AM

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