Sc 634

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-634 is a piece of laboratory equipment designed for scientific research. It is a compact, high-performance centrifuge capable of separating and concentrating various biological samples, such as cells, organelles, and macromolecules. The core function of Sc-634 is to provide efficient and reproducible sample separation through the application of controlled centrifugal forces.

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Lab products found in correlation

4 protocols using sc 634

Intestinal Protein Extraction and Analysis

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Total proteins were extracted from intestine using Pro-Prep™ lysis buffer (Intron Biotechnology, Seoul, Korea) and quantitated with Bradford assay reagent (Bio-Rad, Hercules, CA, USA) following the manufacturer’s protocol. Immunoblots were probed using the following antibodies: VEGF (1:1000, sc-1836, Santa Cruz Biotechnology, CA, USA), eNOS (1:1000, sc-634, Santa Cruz Biotechnology) and β-actin (1:3000, Sigma, MO, USA). The blots were incubated with these antibodies overnight at 4 °C and detected by the luminescence method using ECL solution (NEL104001EA, PerkinElmer, Waltham, MA, USA).

Han N.K., Jeong Y.J., Pyun B.J., Lee Y.J., Kim S.H, & Lee H.J. (2017). Geranylgeranylacetone Ameliorates Intestinal Radiation Toxicity by Preventing Endothelial Cell Dysfunction. International Journal of Molecular Sciences, 18(10), 2103.

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Protein Extraction and Western Blot Analysis

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Total protein was extracted from cell cultures using RIPA lysis buffer (Mbiotech), followed by two washes with DPBS (Welgene) and quantitation via a Bradford assay (Bio-Rad, Hercules, CA, USA) following the manufacturer’s protocol. Equal amounts of total proteins were loaded onto 6–8% SDS–PAGE gels and transferred into NT nitrocellulose membranes (Pall Corporation, Pensacola, FL, USA). Membranes were incubated at 4°C with eNOS (1:1000, sc-634, Santa Cruz Biotechnology, Dallas, TX, USA), p-eNOS (1:1000, #9571s, Cell Signaling Technology, Danvers, MA, USA), p21 (1:1000, sc-6246, Santa Cruz Biotechnology), α-SMA (1:1000, ab5694, Abcam, Cambridge, UK), PECAM-1 (1:1000, ab28364, Abcam), or anti-β-actin (1:3000, #3700, Cell Signaling Technology) in 3% bovine serum albumin (GenDEPOT, Katy, TX, USA). After overnight incubation, the membranes were washed thoroughly in PBS-T buffer and incubated for 1 h with the corresponding secondary antibodies diluted at 1:3000. The proteins were detected using the ECL solution luminescence method (NEL104001EA, PerkinElmer, Waltham, MA, USA).

Lee S.H., Jeong Y.J., Park J., Kim H.Y., Son Y., Kim K.S, & Lee H.J. (2022). Low-Dose Radiation Affects Cardiovascular Disease Risk in Human Aortic Endothelial Cells by Altering Gene Expression under Normal and Diabetic Conditions. International Journal of Molecular Sciences, 23(15), 8577.

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Silencing EPO in ARPE-19 Cells

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siRNA Transfection:
ARPE-19 cells were
seeded in 24-well plates at a density of 6 × 10⁴ cells/well
with DMEM (0.1 mL) containing FBS (10%) and streptomycin/penicillin
(1%). ARPE-19 cells were treated with 175 ng (final concentration
of 23 nM) of siRNA against EPO (Sigma, sense = 5′GACCCUUCAGCUUCAUAUATT,
antisense = 5′UAUAUGAAGCUGAAGGGUCTT) and random sequence control
siRNA (Allstars control negative siRNA, Qiagen, Valencia, CA) in 100
μL of culture medium without serum. Transfection was conducted
with HiPerFect transfection reagent (Qiagen, Valencia, CA), and cells
were harvested 48 h post-transfection. Proteins were assayed by western
blot for detection of EPO (ab226956, Abcam; sc5290, Santa Cruz), AIF
(EMD Millipore), BCL-xL (sc-634, Santa Cruz Biotechnology), caspase-3
(EMD Millipore), BAK (ab32371, Abcam), and actin (sc-47778, Santa
Cruz Biotechnology). Protein localization and expressions were also
analyzed by immunocytochemistry.

Samson F.P., He W., Sripathi S.R., Patrick A.T., Madu J., Chung H., Frost M.C., Jee D., Gutsaeva D.R, & Jahng W.J. (2020). Dual Switch Mechanism of Erythropoietin as an Antiapoptotic and Pro-Angiogenic Determinant in the Retina. ACS Omega, 5(33), 21113-21126.

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Western Blot Analysis of Liver Proteins

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Liver tissue was homogenized in lysis buffer containing 15 mM 3-[(3-cholamidopropyl) dimethyl-ammonio]-1-propanesulfonate, 0.15 M NaCl, 2 mM ethylenediaminetetraacetic acid (pH 8), 1 mM phenylmethanesulfonyl fluoride, 1 mM Na₃VO₄, and Tris-Cl (pH 7.5). Western blot analysis was performed as described previously (17 (link),30 (link)) using primary antibodies specific for goat polyclonal anti-human HB-EGF, goat polyclonal anti-human HGF (AB-294-NA; R&D Systems), mouse monoclonal anti-mouse α-smooth muscle actin (α-SMA; A2547; Sigma-Aldrich), rabbit polyclonal anti-mouse Bcl-2 (sc-492; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit polyclonal anti-mouse Bcl-xL (sc-634; Santa Cruz Biotechnology, Inc.), mouse monoclonal anti-mouse Bax (sc-7480; Santa Cruz Biotechnology, Inc.), mouse monoclonal anti-mouse collagen I (ab88147; Abcam) or mouse monoclonal anti-mouse α-tubulin (T6074; Sigma-Aldrich). The densities of the bands were measured, and the signal ratios of α-SMA/α-tubulin and collagen I/α-tubulin were calculated. For detection of each molecule, 25 µg of protein was loaded (40 µg in the case of α-SMA). Band densities were measured using ImageJ software.

Sakamoto K., Khai N.C., Wang Y., Irie R., Takamatsu H., Matsufuji H, & Kosai K.I. (2016). Heparin-binding epidermal growth factor-like growth factor and hepatocyte growth factor inhibit cholestatic liver injury in mice through different mechanisms. International Journal of Molecular Medicine, 38(6), 1673-1682.

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