dUTP was from Sigma, RNA polymerase III inhibitor (ML-60218) was from Merck Millipore, dNTP (Fermentas), type I IFNα were from PBL Biomedical Laboratories, CpG (Invitrogen), polyIC and peptidoglycan (from Staphylococcus aureus, InvivoGen), Taq Polymerase (BIOTAQ DNA polymerase, Bioline), Pfu (Agilent Technologies), cGAS siRNA (sc-95512), pol III RPC39 siRNA (sc-36292), pol III RPC62 siRNA (sc-76188) were from Santa Cruz Biotechnology, STING (antibody #3337), phospho-IRF-3 (Ser396) (4D4G, rabbit mAb #4947), IRF-3 (D6I4C, XP rabbit mAb #11904), β-catenin antibody (amino-terminal antigen, #9581), mouse anti-rabbit IgG (conformation specific, L27A9, mAb #3678), anti-rabbit IgG, HRP-linked (antibody #7074), cGAS (D1D3G, mAb#15102) antibodies were from Cell Signaling Technology. APOBEC3A antibody (SAB4500753) was from Sigma Aldrich. RIG-I (E-5, sc-376882), pol III RPC39 (C 39-2, sc-23913), pol III RPC62 (I-18, sc-69534) antibodies were from Santa Cruz Biotechnology, Inc. Anti-mouse IgG, (HRP-linked antibody #NA931V) was from GE Healthcare. Monoclonal anti-β-actin−peroxidase (antibody #A3854) was from Sigma Aldrich. Human IFNα (IgA, mba-hifnα-3), hIFNβ (IgG, mbg-hifnβ-3), hIFNγ (IgA, mba-hifnγ-3) and Mouse IgG2a (mbg2a-ctrlm) antibodies used as control were from InVivogen.
Cgas sirna sc 95512
Manufactured by Santa Cruz Biotechnology
CGAS siRNA (sc-95512) is a product offered by Santa Cruz Biotechnology. It is a small interfering RNA (siRNA) designed to target and silence the expression of the CGAS (cyclic GMP-AMP synthase) gene in cellular systems.
Automatically generated - may contain errors
Lab products found in correlation
β catenin antibody, by Cell Signaling Technology (1 mentions)
Rabbit anti mouse igg, by Cell Signaling Technology (1 mentions)
Cgas d1d3g, by Cell Signaling Technology (1 mentions)
Monoclonal anti β actin peroxidase, by Merck Group (1 mentions)
Sc 36868, by Santa Cruz Biotechnology (1 mentions)
Sc 29528, by Santa Cruz Biotechnology (1 mentions)
2 protocols using cgas sirna sc 95512
1
Dissecting cGAS-STING Signaling Pathway
2
siRNA Transfection and DNA Transfection
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The siRNA transfection was performed according to the manufacturer’s protocol. In total, 2 × 105 cells per well were seeded in DMEM media with supplements in a six-well dish and incubated overnight; media was replaced with DMEM without antibiotics 4 h ahead of the transfection. First, 20 pmol siRNA (cGAS siRNA sc-95512, CTRL siRNA sc-36869, Santa Cruz) was diluted in 100 µL of transfection medium (SC-36868, Santa Cruz) and 6 µL of transfection reagent (SC-29528, Santa Cruz) were diluted to 100 µL with transfection medium. Then, siRNA and transfection reagent solution were gently mixed and incubated at room temperature for 30 min. Next, the entire 200-µL siRNA/transfection solution was added, dropwise, on top of cells. Cells were then left to incubate for 16 h. The next day, the media was exchanged for fully supplemented media, and the cells were incubated for 24 h before collection for RT-qPCR, Western blot, or immunofluorescence. Double- stranded ISD DNA oligo (10 μg) and plasmid DNA were transfected with a similar strategy and collected after 4 and 6 h for qPCR analysis.
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