Socs 1 sirna

Manufactured by Santa Cruz Biotechnology

Sourced in United States

SOCS-1 siRNA is a small interfering RNA molecule designed to target the SOCS-1 gene. SOCS-1 is a negative regulator of cytokine signaling pathways. The siRNA is intended for use in cellular and molecular biology research applications to modulate SOCS-1 expression and study its functions.

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4 protocols using socs 1 sirna

Artesunate Anticancer Signaling Pathway

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Artesunate (ART), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), Tris base, glycine, NaCl, sodium dodecyl sulfate (SDS), RNase A, DPX mountant for histology, and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO). Iscove Modified Dulbecco Medium (IMDM), RPMI 1640, and fetal bovine serum (FBS) were obtained from Lonza Group Ltd. (Basel, Switzerland). 0.4% Trypan Blue solution, and antibiotic-antimycotic mixture was obtained from Life Technologies (Grand Island, NY). Anti-phospho-p38, anti-p38, anti-phospho-ERK, anti-ERK, anti-phospho-CREB, anti-CREB, anti-phospho-JAK2, anti-JAK2, anti-procaspase-3, and anti-cleaved caspase-3 antibodies were purchased from Cell Signaling Technology (Beverly, MA). Anti-phospho-STAT5, anti-STAT5, anti-SOCS-1, SOCS-1 siRNA, anti-bcl-2, anti-bcl-xL, anti-survivin, anti-cyclin D1, anti-IAP-1, anti-IAP-2, anti-PARP, anti-Ki-67, anti-VEGF, anti-β-actin, and horseradish peroxidase (HRP)-conjugated secondary antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). TUNEL (terminal transferase mediated dUTP-fluorescein nick end labeling) assay kit was from Roche Diagnostics GmbH (Mannheim, Germany). Whole-cell lysates of tumor tissues were obtained with T-PER Tissue Protein Extraction Reagent (Pierce, Rockford, USA).

Kim C., Lee J.H., Kim S.H., Sethi G, & Ahn K.S. (2015). Artesunate suppresses tumor growth and induces apoptosis through the modulation of multiple oncogenic cascades in a chronic myeloid leukemia xenograft mouse model. Oncotarget, 6(6), 4020-4035.

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SOCS1 Silencing Impacts Cellular Responses

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The SOCS1-siRNA and scrambled-siRNA (SC-siRNA) were purchased from Santa Cruz Biotechnology (USA). The cells were transfected with 60 pmol SOCS1-siRNA using the Lipofectamine reagent (Invitrogen, USA) in serum-free medium according to the manufacturer's instructions. The cells were incubated for 6 h and recovered for an additional 6 h before the treatments. The experiments were repeated four times (n = 4). The SC-siRNA served as the negative control.

Cai H., Liang Q, & Ge G. (2016). Gypenoside Attenuates β Amyloid-Induced Inflammation in N9 Microglial Cells via SOCS1 Signaling. Neural Plasticity, 2016, 6362707.

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Microglial Cell Activation and Regulation

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N9 cell, a mouse microglial cell line, was obtained from the Fourth Military Medical University, China. The N9 microglial cells are very similar to the primary cultured microglial cells in producing cytokines and reacting to stimulus. The IMDM cell culture medium, fetal bovine serum (FBS), LPS (Escherichia coli, O55:B5), IFNγ, methylthiazol tetrazolium (MTT), and resveratrol were purchased from Sigma-Aldrich (USA). The SIRT1-siRNA, SOCS1-siRNA, and scrambled- (SC-) siRNA were purchased from Santa Cruz Biotechnology (USA). Antibodies against iNOS, SIRT1, SOCS1, and NF-κB p65 subunit were purchased from the Abcam (Cambridge, UK). Antibodies against β-actin and GAPDH and Cy3-labeled secondary antibody were obtained from the Beijing Comwin Biotech Co., Ltd. (China).

Zhang S., Gao L., Liu X., Lu T., Xie C, & Jia J. (2017). Resveratrol Attenuates Microglial Activation via SIRT1-SOCS1 Pathway. Evidence-based Complementary and Alternative Medicine : eCAM, 2017, 8791832.

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Efficient Electroporation for siRNA Delivery in KBM-5 Cells

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We investigated the ability of commercially available electroporation systems, the Neon™ Transfection System (Invitrogen, Carlsbad, CA). Transfection efficiency was measured by Western blot analysis. KBM-5 cells were prepared for transfection, after cells were resuspended with 120 μl of Neon Resuspension Buffer R for every one million cells. For each electroporation, KBM-5 cells with 50 nM of SOCS-1 siRNA (Santa Cruz, CA) were aliquoted into a sterile microcentrifuge tube. A Neon Tip was inserted into the Neon Pipette and the cell-siRNA mixture was aspirated into the tip avoiding air bubbles. The Neon Pipette was then inserted into the Neon Tube containing 3 ml of Neon Electrolytic Buffer E in the Neon Pipette Station. Cells were pulsed once with a voltage of 1,300 and a width of 20 ms. After 48 h of transfection, cells were treated with 100 μM of ART for 4 h, and whole-cell extracts were washed twice with ice-cold PBS, lysed with lysis solution, and cell lysates were prepared for Western blot analysis.

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