Sc 29471 5

Manufactured by Santa Cruz Biotechnology

Sc-29471-V is a laboratory product manufactured by Santa Cruz Biotechnology. It is a research-grade tool designed for use in scientific experiments and investigations. The core function of this product is to facilitate specific research activities, though detailed specifications are not available at this time.

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Lab products found in correlation

Protein assay, by Bio-Rad (1 mentions) Lambda phosphatase, by Santa Cruz Biotechnology (1 mentions) Ripa buffer, by Merck Group (1 mentions) Complete protease inhibitor cocktail, by Roche (1 mentions) Mkn45, by American Type Culture Collection (1 mentions)

3 protocols using sc 29471 5

Investigating RhoA Phosphorylation in Gastric Cancer

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AGS is a Lauren intestinal-type gastric adenocarcinoma cell line, and MKN-45 is a Lauren diffuse-type gastric adenocarcinoma cell line. AGS and MKN-45 cells were obtained from the America Type Culture Collection (ATCC). Cell lysates were collected in RIPA buffer (Sigma) containing Complete Protease Inhibitor Cocktail (Roche Diagnostics), and protein concentration was determined by the Bio-Rad Protein Assay. Anti-phosphorylated( Ser188)-RhoA antibody (Abcam ab125275, Cambridge, MA) was used to report phosphorylated RhoA (p-RhoA), and anti-RhoA (67B9) antibody (Cell Signaling #2117, Danvers, MA) was used to report total RhoA expression.
AGS and MKN-45 cell lysates were treated with lambda phosphatase (Santa Cruz Biotechnology sc-200312, Dallas, TX) to abolish immunoreactivity to phospho-dependent antibodies.
Silencing of RhoA was achieved via lentiviral transduction of human RhoA shRNA (sc-29471-V; Santa Cruz Biotechnology). A scramble shRNA control (SC-108080) was also used.

Chang K.K., Cho S.J., Yoon C., Lee J.H., Park D.J, & Yoon S.S. (2016). Increased RhoA Activity Predicts Worse Overall Survival in Patients Undergoing Surgical Resection for Lauren Diffuse-Type Gastric Adenocarcinoma. Annals of surgical oncology, 23(13), 4238-4246.

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Efficient Rho A Knockdown in MG63 Cells

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Knockdown of Rho A was achieved via lentiviral transduction of human Rho A shRNA (sc-29471-V; Santa Cruz Biotechnology). According to the manufacturer’s instructions, scrambled shRNA (SC-108080) and the mCherry control (LPP-MCHR-LV105-025-C) were also included. The lentiviral particle solution was added to MG63 cells in fresh DMEM containing 5 mg/mL polybrene. The medium was changed after 12 h of infection. After successive 12-h intervals, the medium was changed to fresh DMEM, and the cells were cultured for 72 to 96 h. After transduction, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis was employed to determine the knockdown efficiency.

Wang J., Zhang L., Qu R., Zhang L, & Huang W. (2018). Rho A Regulates Epidermal Growth Factor-Induced Human Osteosarcoma MG63 Cell Migration. International Journal of Molecular Sciences, 19(5), 1437.

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RhoA Gene Silencing via Lentiviral Transduction

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Silencing of RhoA was achieved via lentiviral transduction of human RhoA shRNA (sc-29471-V; Santa Cruz Biotechnology). A scramble shRNA control (SC-108080) and a GFP control (sc108084) were also used. Maximal knockdown of RhoA occurred 72 to 96 hours after transduction.

Yoon C., Cho S.J., Aksoy B.A., Park D.J., Schultz N., Ryeom S.W, & Yoon S.S. (2015). Chemotherapy resistance in diffuse type gastric adenocarcinoma is mediated by RhoA activation in cancer stem-like cells. Clinical cancer research : an official journal of the American Association for Cancer Research, 22(4), 971-983.

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