Table 2.
Antibodies used in Western blot and IF studies.
| Target | Antibody | Catalog Nr. | Supplier |
|---|---|---|---|
| Primary | |||
| HIF-1α (WB) | Rabbit polyclonal | #3716 | Cell Signaling Technology, Leiden, The Netherlands |
| HIF-1α (IF) | Rabbit monoclonal | ab51608 | Abcam, Cambridge, UK |
| Pro-OH HIF-1α (WB) | Rabbit monoclonal | 3434 | Cell Signaling Technology, Danvers, MA, USA |
| PHD2/EGLN1 (IP-WB) | Rabbit polyclonal | NB100-137 | Novus Biologicals, Centennials, CO, USA |
| HK2 (WB) | Mouse monoclonal | ab104836 | Abcam, Cambridge, UK |
| PKM2 (WB) | Rabbit polyclonal | #3198 | Cell Signaling Technology, Danvers, MA, USA |
| LDHA (WB) | Rabbit polyclonal | 19987-1-AP | Proteintech, Manchester, UK |
| PDK1 (WB) | Rabbit monoclonal | #3820 | Cell Signaling Technology, Danvers, MA, USA |
| MT-CO1 (IF) | Mouse monoclonal conjugated to Alexa Fluor® 488 | Ab154477 | Abcam, Cambridge, UK |
| MGO (IP-WB) | Mouse monoclonal | NBP-2 59368 | Novus Biologicals, Centennials, CO, USA |
| IgG2 Isotype control (IP) | Mouse monoclonal | ab18415 | Abcam, Cambridge, UK |
| IgG Isotype control (IP) | Rabbit monoclonal | Ab172730 | Abcam, Cambridge, UK |
| Β-actin | Mouse monoclonal | A5441 | Sigma Aldrich, St. Louis, MO, USA |
| Histone H3 (WB) | Rabbit polyclonal | ab1791 | Abcam, Cambridge, UK |
| Secondary | |||
| HIF-1α (WB), Pro-OH HIF-1α, PHD2/EGLN1, PKM2, LDHA, PDK1 | HRP-conjugated goat anti-rabbit | P0448 | Agilent/Dako, Santa Clara, CA, USA |
| B-actin, HK2, MGO | HRP-conjugated goat anti-mouse | P0447 | Agilent/Dako, Santa Clara, CA, USA |
| HIF-1α (IF) | Alexa Fluor® Plus 488 goat anti-rabbit IgG | A-32731 | Thermo Fisher Scientific, Waltham, MA, USA |
HIF-1α = hypoxia inducible factor 1α; WB = Western blot; IF = immunofluorescence; Pro-OH HIF-1α = proline-hydroxylated hypoxia-inducible factor 1α; PHD2/EGLN1 = prolyl hydroxylase domain-containing protein 2/egl-9 family hypoxia inducible factor 1; HK2 = hexokinase 2; PKM2 = pyruvate kinase isozymes M2; LDHA = lactate dehydrogenase A; PDK1 = pyruvate dehydrogenase kinase 1; MT-CO1 = mitochondrially encoded cytochrome C oxidase I; MGO = methylglyoxal; IP = immunoprecipitation.