Sc 8418

Manufactured by Santa Cruz Biotechnology

Sourced in United Kingdom, United States

Sc-8418 is a laboratory instrument used for protein expression and purification. It is designed to facilitate the isolation and extraction of target proteins from biological samples.

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Lab products found in correlation

Vectashield with dapi, by Vector Laboratories (1 mentions) Metamorph, by Universal Imaging (1 mentions) Ab18197, by Abcam (1 mentions) Ab98023, by Abcam (1 mentions) Ab8805, by Abcam (1 mentions) Anti β catenin sc 7963, by Santa Cruz Biotechnology (1 mentions) Bca protein assay kit, by Bioss Antibodies (1 mentions) Sc 516102 sc 2357, by Santa Cruz Biotechnology (1 mentions) Rpmi 1640 medium, by Yuanye Bio-Technology (1 mentions) T per lysis buffer, by Thermo Fisher Scientific (1 mentions) C digit blot scanner, by LI COR (1 mentions) Anti actin, by Merck Group (1 mentions) Goat anti mouse igg horseradish peroxidase, by Jackson ImmunoResearch (1 mentions) A4700, by Merck Group (1 mentions) Anti phospho s6, by Cell Signaling Technology (1 mentions) Phosphatase inhibitor, by Roche (1 mentions) Ab84422, by Abcam (1 mentions) Ab51081, by Abcam (1 mentions) Orca r2 cooled ccd camera, by Hamamatsu Photonics (1 mentions) Ix81 widefield microscope, by Hamamatsu Photonics (1 mentions)

5 protocols using sc 8418

Imaging of P-bodies in Fixed Cells

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Fixed cells were imaged at room temperature on an Axioimager Z1 wide-field microscope (63×, NA 1.4; ZEISS) equipped with an sCMOs Zyla 4 0.2 camera (Andor Technology) and controlled by MetaMorph (Universal Imaging). 3D image stacks were collected with a Z-spacing of 0.3 µm. Figures were prepared with ImageJ (National Institutes of Health), Photoshop (Adobe Systems), and Illustrator (Adobe Systems), and graphs were generated with R.
P-bodies were labeled with a mouse monoclonal antibody raised against S6K and that is known to recognize GE-1/Helds (sc-8418; Santa Cruz Biotechnology, Inc.; Kedersha and Anderson, 2007 (link)). The secondary anti-mouse antibody was coupled to FITC. Slides were mounted in Vectashield with DAPI (Vector Laboratories).

Pichon X., Bastide A., Safieddine A., Chouaib R., Samacoits A., Basyuk E., Peter M., Mueller F, & Bertrand E. (2016). Visualization of single endogenous polysomes reveals the dynamics of translation in live human cells. The Journal of Cell Biology, 214(6), 769-781.

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Protosappanin B Anti-Cancer Protocol

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Protosappanin B was purchased from Shanghai Yuanye Bio-Technology Co. Ltd. (B21622-20 mg, Shanghai, China). Reagents, signaling pathway agonists and antibodies were purchased from the following companies: RPMI-1640 medium, DMEM medium (Shanghai Yuanye Bio-technology, Shanghai, China); Streptavidin Peroxidase immunohistochemistry kit (GS4961, GS4962), cell counting kit-8 (QN1293), Annexin V-FITC/PI double staining apoptosis detection kit (SNM530), and ECL western blotting detection kit (QN1155) (Beijing BioRab Technology Co. Ltd., Beijing, China); BCA protein assay kit (C05-02001), western blocking buffer (5% BSA, C05-06002) (Beijing Biosynthesis Biotechnology Co., Ltd, Beijing, China); IGF-1 (K002504P), AZD2858 (A126819), and TPA (bs-1545R) (Shanghai Hengfei Biotechnology Co. Ltd. Shanghai, China); Anti-GOLPH3 (ab98023), Anti-p-AKT (ab8805), Anti-AKT (ab38449), and anti-PCNA (proliferating cell nuclear antigen) (ab18197) (Abcam, Cambridge, UK); Anti-p-p70S6K (ribosomal protein S6 kinase, 70 kDa) (sc-8416), Anti-p70S6K (sc-8418), Anti-β-catenin (sc-7963), Anti-p-ERK1/2(extracellular signal-regulated kinase 1 and 2) (sc-81492), Anti-ERK1/2 (sc-514302), Anti-GAPDH (sc-365062), and secondary antibodies (sc-516102, sc-2357) (Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Zheng X.C., Shi Z.S., Qiu C.Z., Hong Z.S., Wang C.X., Zhuang H.B., Chen Z.C, & Pan J.P. (2020). Protosappanin B Exerts Anti-tumor Effects on Colon Cancer Cells via Inhibiting GOLPH3 Expression. Integrative Cancer Therapies, 19, 1534735420972477.

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Kidney Protein Analysis by Western Blot

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Kidneys were collected and homogenized in T-PER lysis buffer (Thermo Fisher Scientific) supplemented with protease (Pierce) and phosphatase inhibitors (Roche). The homogenized lysate was incubated at 4°C for 20 min and cell debris cleared by centrifugation at 12,000 g for 10 min. 50 µg of protein lysate from each kidney was analyzed by SDS-PAGE. Membranes were incubated overnight at 4°C with the following primary antibodies: anti–phospho–p-S6K (1:200; Tyr 389; 9205; Cell Signaling Technology), anti–p-S6K (1:1,000; sc-8418; Santa Cruz Biotechnology), anti–phospho-S6 (1:200; Ser235/236; 2211; Cell Signaling Technology), anti-S6 (1:500; sc-74459; Santa Cruz Biotechnology), and anti-actin (1:1,000; A4700; Sigma-Aldrich). Secondary antibodies included donkey anti–rabbit IgG–horseradish peroxidase (Sigma-Aldrich) and goat anti–mouse IgG–horseradish peroxidase (Jackson ImmunoResearch Laboratories). Blots were imaged on a C-DiGit Blot Scanner (LI-COR), and signal intensities for each protein were quantified and normalized to actin.

Dionne L.K., Shim K., Hoshi M., Cheng T., Wang J., Marthiens V., Knoten A., Basto R., Jain S, & Mahjoub M.R. (2018). Centrosome amplification disrupts renal development and causes cystogenesis. The Journal of Cell Biology, 217(7), 2485-2501.

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Multicolor Immunofluorescence Staining Protocol

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Cells were fixed for 5 min in 3.7% paraformaldehyde (PFA) (Sigma-Aldrich) followed by 2 min permeabilization with 0.5% NP-40 (EMD Millipore). The coverslips were incubated with blocking solution (1% bovine serum albumin) for 1 h at RT. Primary antibodies used for immune staining were diluted in blocking solution and applied to the coverslips for 1 h at RT; this was followed by three short rinses in PBS. Primary antibodies were as follows: anti-Matr3C (ab84422; Abcam), anti-Matr3N (ab51081; Abcam), and mouse anti-S6K used to detected Hedls (sc-8418; Santa Cruz) at 1:200; rabbit anti-Rck/p54 (A300-461A; Bethyl Laboratories, Montgomery, TX) and mouse anti-PABP-1 (10E10, sc-32318; Santa Cruz) at 1:200. Next the coverslips were incubated with secondary antibodies, diluted 1:1000 in blocking buffer, and conjugated to Alexa Fluor 405, 488, or 546 (Invitrogen, Carlsbad, CA) for 1 h at RT. The coverslips were next stained with 4′,6-diamidino-2-phenylindole and rinsed with PBS before being mounted onto slides using Mowiol mounting media. Images were acquired at RT using a using an Olympus IX81 wide-field microscope with a 40× air objective lens attached to a Hamamatsu Orca-R2 cooled CCD camera. Pearson’s colocalization coefficients were determined using the Coloc2 plug-in in imageJ.

Rajgor D., Hanley J.G, & Shanahan C.M. (2016). Identification of novel nesprin-1 binding partners and cytoplasmic matrin-3 in processing bodies. Molecular Biology of the Cell, 27(24), 3894-3902.

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Antibody Immunostaining Assay Protocol

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Antibodies against p70S6K (sc-8418), LAMP1 (sc-19992), and phospho-mTOR (sc-293133) were purchased from Santa Cruz Biotechnology (USA). Antibodies against phosphor-p70S6K (AP0564), RRAGA (A15134), RRAGD (A9979), and ADORA1 (A5219), ADORA2A (A1587) were purchased from ABclonal (China). Antibodies against NT5C2 (15223-1-AP), RRAGB (13023-1-AP), RRAGC (26989-1-AP), SP1 (21962-1-AP), YY1 (66281-1-lg), mTOR (66888-1-Ig), Actin (66009-1-lg), GAPDH (10494-1-AP), ADA1 (13328-1-AP), MDH2 (15462-1-AP), MDH1 (15904-1-AP), ACO1 (12406-1-AP), CS (16131-1-AP), GLUD1 (14299-1-AP), and SDHB (10620-1-AP) were purchased from Proteintech (China). Antibody against PNP (DF8260) was purchased from Affinity (China). Antibody against puromycin (MABE343) was purchased from Sigma-Aldrich (USA). Antibody against Ki67 (ab15580) was purchased from Abcam (UK).

Li M.X., Wu X.T., Jing W.Q., Hou W.K., Hu S, & Yan W. (2023). Inosine enhances tumor mitochondrial respiration by inducing Rag GTPases and nascent protein synthesis under nutrient starvation. Cell Death & Disease, 14(8), 492.

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