Hif 1β

Manufactured by Santa Cruz Biotechnology

Sourced in United States

HIF-1β is a subunit of the hypoxia-inducible factor 1 (HIF-1) complex, a key transcriptional regulator that responds to changes in available oxygen in the cellular environment. HIF-1β is a basic helix-loop-helix (bHLH) protein that dimerizes with the HIF-1α subunit to form the active HIF-1 complex.

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Lab products found in correlation

4 protocols using hif 1β

Western Blot Analysis of Gut Markers

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Proteins were separated by SDS gels and transferred to polyvinylidene difluoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA). The membranes were blocked with 3% bovine serum albumin prepared in TBS (Tris-buffered saline, pH 7.5 containing 0.1% Tween-20) for 1 h and then incubated with antibodies overnight at 4 °C: anti-p65 (1:200. Santa Cruz, CA, USA), anti-IκBα (1:200. Santa Cruz, CA, USA), IKKβ (1:200. Santa Cruz, CA, USA), mucin-2 (Muc-2, 1:500. Abcam, Cambridge, MA, USA), intestinal trefoil factor (ITF, 1:500. Abcam, Cambridge, MA, USA), ZO-1 (1:500. Abcam, Cambridge, MA, USA), Occludin (1:500. Abcam, Cambridge, MA, USA), HIF-1α (1:500. Santa Cruz, CA, USA), HIF-1β (1:500. Santa Cruz, CA, USA), Cleaved-caspase-3 (1:500. Santa Cruz, CA, USA), Claudin-1 (1:500. Abcam, Cambridge, MA, USA), and anti-GAPDH (1:1000. Santa Cruz, CA, USA). The membranes were then washed three times in TBST (50 mM Tris-HCl pH 7.5, 140 mM NaCl, 0.1% Tween) and incubated with secondary antibody at room temperature for 1 h. An enhanced chemiluminescence reagent, ECL Western blotting detection reagent (Tanon, Shanghai, China), was used to make the labeled protein bands detectable with Image System Minichemi (Saizhi, Beijing, China).

Wu C., Wang X., Jiang T., Li C., Zhang L., Gao X., Tian F., Li N, & Li J. (2016). Partial Enteral Nutrition Mitigated Ischemia/Reperfusion-Induced Damage of Rat Small Intestinal Barrier. Nutrients, 8(8), 502.

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Panaxotriol Modulates Hypoxia Signaling

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20(R)-panaxotriol was obtained from Nanjing Dilger Medical Technology Co.Ltd. All organic reagents used for the experiment were analytically pure, and other chemicals were purchased from Aladdin reagent. Hep3B cells and HeLa cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% heat-inactivated fetal bovine serum (FBS, Hyclone, Logan, UT, USA), penicillin (100 U/mL), and streptomycin (100 U/mL) (Invitrogen, Carlsbad, CA, USA). The hypoxic culture was kept in a gas-controlled chamber (Thermo Electron Corp., Marietta, OH, USA) and maintained at 37 °C under 5% CO₂, 94% N₂, and 1% O₂. Antibodies against the hypoxia-inducible factor (HIF)-1α were purchased from BD Biosciences (San Diego, CA, USA). Antibodies against the vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-9, HIF-1β, B cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and topoisomerase (Topo)-I were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against cleaved Caspase-3 were purchased from Cell Signaling Technology. Other cell culture reagents were obtained from commercial companies (UNIV, Shanghai, China; Huayi Biotechnology Co., Ltd, Changchun, China).

Guo H.Y., Xing Y., Sun Y.Q., Liu C., Xu Q., Shang F.F., Zhang R.H., Jin X.J., Chen F., Lee J.J., Kang D., Shen Q.K, & Quan Z.S. (2022). Ginsengenin derivatives synthesized from 20(R)-panaxotriol: Synthesis, characterization, and antitumor activity targeting HIF-1 pathway. Journal of Ginseng Research, 46(6), 738-749.

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ChIP-seq Protocol for HIF-1 Factors

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MHCC-97L cells were crosslinked with formaldehyde, lysed with SDS buffer and sonicated. Sheared DNA was precleared with salmon sperm DNA/protein A agarose slurry (Merck Millipore) and immunoprecipitated with HIF-1α, HIF-1β, and IgG (Santa Cruz). Agarose beads were incubated with antibody/protein/DNA complex and washed with low-salt buffer, high-salt buffer, and LiCl wash buffer according to the manufacturer’s protocol (Millipore). DNA was eluted in 1% SDS/0.1 M NaHCO₃ and de-crosslinked with 0.2 M NaCl.

Chiu D.K., Tse A.P., Xu I.M., Di Cui J., Lai R.K., Li L.L., Koh H.Y., Tsang F.H., Wei L.L., Wong C.M., Ng I.O, & Wong C.C. (2017). Hypoxia inducible factor HIF-1 promotes myeloid-derived suppressor cells accumulation through ENTPD2/CD39L1 in hepatocellular carcinoma. Nature Communications, 8, 517.

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Immunoblotting of Cancer Cell Signaling

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OVCAR-3 cancer cells were seeded and incubated overnight before treatment with GA. After a double wash with cold PBS, the cells were harvested with M-PER Mammalian Protein Extraction reagent supplemented with Halt protease and phosphatase inhibitor (both from Pierce), and total protein levels were assayed with the BCA protein assay kit. Cell lysates (50 μg total protein) were separated by SDS-PAGE and blotted onto a nitrocellulose membrane with a Mini-Protean 3 system (Bio-Rad, Hercules, CA, USA). For immunodetection, antibodies against HIF-1α, HIF-1β, AKT, p-AKT, PTEN and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were applied, and signals were visualized with phycoerythrin-conjugated anti-mouse IgG secondary antibodies, SuperSignal West Pico Substrate, and X-ray film (Pierce). Protein bands were quantitated with NIH ImageJ software and normalized by GAPDH bands for analysis.

HE Z., CHEN A.Y., ROJANASAKUL Y., RANKIN G.O, & CHEN Y.C. (2015). Gallic acid, a phenolic compound, exerts anti-angiogenic effects via the PTEN/AKT/HIF-1α/VEGF signaling pathway in ovarian cancer cells. Oncology Reports, 35(1), 291-297.

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