Epas 1

Manufactured by Santa Cruz Biotechnology

EPAS-1 is a laboratory product offered by Santa Cruz Biotechnology. It is an enzyme that plays a role in the regulation of cellular responses to changes in oxygen levels. The core function of EPAS-1 is to act as a transcription factor that activates the expression of genes involved in the adaptive response to hypoxia, or low oxygen conditions.

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Lab products found in correlation

2 protocols using epas 1

Hypoxia and cAMP Regulation in Cells

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Cells were cultured in HAM’s‐F12 medium containing 10% (v/v) FCS without antibiotics for 48 h prior to transfection by Nucleofection (Amaxa Biosystems, Germany). Following trypsinzation 2 × 10⁶ cells were resuspended in 100 µl of cell line solution L with either 300nM of control siRNA‐A (Santa Cruz sc‐37007) or targeted si‐RNA EPAS‐1 (Santa Cruz sc‐35316) and transfected following the manufacturers' instructions using programs X‐005 as previously detailed.16 Following transfection, cells were seeded at approximately 5 × 10⁵/well of a 6‐well plate in HAM’s‐F12 medium containing 10% (v/v) FCS without antibiotics. After 24 hours, the medium was replaced with DMEM/HAM’s F‐12 containing 10% (v/v) FCS and incubated for 48 h in 1% O₂ with and without 100 µM 8Br‐cAMP.

Abid N., Embola J., Tryfonos Z., Bercher J., Ashton S.V., Khalil A., Thilaganathan B., Cartwright J.E, & Whitley G.S. (2020). Regulation of stanniocalcin‐1 secretion by BeWo cells and first trimester human placental tissue from normal pregnancies and those at increased risk of developing preeclampsia. The FASEB Journal, 34(5), 6086-6098.

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Western Blot Analysis of Stem Cell Markers

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Total cell lysates were obtained by adding RIPA buffer (R0728, Sigma-Aldrich) and protease inhibitor cocktail (P2714, Sigma-Aldrich). Equilibrated protein samples were loaded into a 10% SDS-Page gel, electrophoresed and then electro-transferred to a PVDF membrane (Bio-Rad). Transferred membranes were blocked with a blocking buffer containing 5% non-fat milk (Labscientific, M0841). The following monoclonal antibodies were used: Nanog (Cell Signaling, D73G4), CD133 (Cell Signaling, D2V8Q), CD44 (Cell Signaling, 8E2), ALDH1/2 (Santa Cruz, sc-166362), ALDH1 (Cell Signaling, D9J7R), MDR1/ABCB1 (Cell Signaling, E1Y7B), HIF1-α (Cell Signaling, D5F3M), ABCG2 (Cell Signaling, D5V2K), nucleoporin-62 (Santa Cruz, sc-48373), EpCAM (Cell Signaling, VU1D9) and EPAS-1 (Santa Cruz, 190B). Target proteins were visualized with the secondary mouse or rabbit IgG antibodies and a chemiluminescent substrate (Santa Cruz, sc-2048). All Western blot results were normalized by total protein (Fig. S5). All Western blot experiments were reproduced and performed three times for quantification.

Shen Q., Hill T., Cai X., Bui L., Barakat R., Hills E., Almugaiteeb T., Babu A., Mckernan P.H., Zalles M., Battiste J.D, & Kim Y.T. (2021). Physical Confinement during Cancer Cell Migration Triggers Therapeutic Resistance and Cancer Stem Cell-like Behavior. Cancer letters, 506, 142-151.

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