Sc 7993

Manufactured by Santa Cruz Biotechnology

Sc-7993 is a laboratory equipment product offered by Santa Cruz Biotechnology. The core function of Sc-7993 is to [description not available].

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Lab products found in correlation

3 protocols using sc 7993

Immunostaining of HP1α and H3K9me3

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HeLa cells were plated onto 6 well plates at a concentration of 0.3×10⁶ cells/well and allowed to adhere overnight. The next day cells were treated with 10 uM streptonigrin for 6 hours. Afterwards cells were fixed with 500 ul/well 3.7% (v/v) formaldehyde for 15 minutes, permeabilized with 500 μl of 0.3% PBT and 1% BSA in PBS for 20 minutes. 500 ul of primary antibody against HP1α (or CBX5) (1:200; Life Sciences, 730019) was then spread and allowed to incubate overnight at 4 °C. The next day, cells were then incubated with secondary antibody at room temperature for 120 minutes, and mounted on coverslip with DAPI and observed under fluorescence microscope.
Primary antibodies used in immunostaining and Western blotting include mouse antibodies against HP1α (or CBX5) (1:200; Life Sciences, 730019), rabbit antibodies against H3K9me3 (1:500, MilliporeSigma (Burlington, MA), 07–442), goat anti-p-STAT3 (1:250, Santa Cruz, sc-7993), and rabbit anti-STAT3 (1:500; Santa Cruz, sc-482).

Loyola A.C., Dao K., Shang R., Zhang L., Dutta P., Fowler C., Li J, & Li W.X. (2020). Streptonigrin at low concentration promotes heterochromatin formation. Scientific Reports, 10, 3478.

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Pluripotency Protein Expression Analysis

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Whole-cell protein extracts were isolated from human cells with conventional RIPA lysis buffer and protein concentration assayed by BCA kit (Thermo Scientific). Blocking was carried out in 10% skim milk in PBST for 1 h. Blots were incubated with the following antibodies in 5% BSA in PBST: KLF4 (AF3158; 1:200; R&D), OCT4 (H-134; 1:1,000; Santa Cruz), NANOG (397A; 1:1,000; Bethyl), METTL3 (A301-567A, 1:2000, Bethyl), HSP90beta (ab32568, 1:10000, Abcam), DNMT1 (ab87654, 1:1000, Abcam), GAPDH (ab181602, 1:10000, Abcam), ACTIN (ab6276, 1:10000, Abcam), UHRF1 (sc-373750, 1:1000, Santa Cruz), DGCR8 (10996-1-AP, 1:1000, Proteintech), P53 (D-01, courtesy from Varda Roter’s lab), β-catenin (sc-7963, 1:1000, Santa Cruz), TCF3 (CST2883, 1:1000, Cell Signaling), STAT3 (sc-482, 1:1000, Santa Cruz), KLF4 (sc-20691, 1:1000, Santa Cruz), RBPJ (C5 5313, 1:1000, Cell Signaling), TFAP2C (CSTH2320, 1:1000, Cell Signaling), STAT3 (sc-7993, 1:1000, Santa Cruz). Secondary antibodies were HRP-linked goat anti-mouse, goat anti-rabbit and rabbit anti-goat (1:10,000; Jackson). Blots were developed using SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific 34580).

Bayerl J., Ayyash M., Shani T., Manor Y.S., Gafni O., Massarwa R., Kalma Y., Aguilera-Castrejon A., Zerbib M., Amir H., Sheban D., Geula S., Mor N., Weinberger L., Naveh Tassa S., Krupalnik V., Oldak B., Livnat N., Tarazi S., Tawil S., Wildschutz E., Ashouokhi S., Lasman L., Rotter V., Hanna S., Ben-Yosef D., Novershtern N., Viukov S, & Hanna J.H. (2021). Principles of signaling pathway modulation for enhancing human naive pluripotency induction. Cell Stem Cell, 28(9), 1549-1565.e12.

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Western Blot Analysis of RTHy Cells

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The supernatants of RTHy cell extracts in SDS-PAGE loading buffer were heated at 95°C for 6 min. The prepared samples were separated in SDS-PAGE gel, with 10 µL of loading for the analysis of α-tubulin and 20 µL for the analysis of Stat3, pStat3, and pAkt. The separated proteins were transferred onto a PVDF membrane (Bio-Rad), and blocked in 5% (w/v) skimmed milk, 50 mmol L -1 Tris-HCl, pH 7.4, and 0.05% (v/v) Tween-20 (TBST) for 2 h. The antibodies against α-tubulin (1:2000 dilution; T9026, Sigma-Aldrich), Stat3, phosphor-Stat3 (Tyr705) and phospho-Akt1/2/3 (Ser473) (1:200 dilution; sc-7179, sc-7993 and sc-7985, Santa Cruz Biotechnology) were diluted in 3% (w/v) BSA, TBST, and incubated with the PVDF membranes overnight at 4°C. The membranes were washed with TBST for 6 × 5 min, and then incubated with anti-mouse/rabbit IgG horseradish peroxidase-linked antibody (1:33,000 dilution; GE Healthcare) and diluted in 3% BSA, TBST for 1 h. Then the membranes were washed with TBST for 6 × 10 min. The antibody-bound protein bands were detected by ECL plus western-blotting detection system and imaged on high performance chemiluminescence film (GE Healthcare).

Gong N., Jönsson E, & Björnsson B.T. (2016). Acute anorexigenic action of leptin in rainbow trout is mediated by the hypothalamic Pi3k pathway. Journal of molecular endocrinology, 56(3).

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