Stat1α p91 Antibody (C-111): sc-417

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

Whole cell extracts were separated on Mini Protean TGX gels and transferred onto nitrocellulose membrane (BioRad®). SINV-specific ascites fluid (ATCC-VR-1248AF) and antibodies specific to actin (Thermo Fisher®, MS-1295-P), GAPDH (Sigma®, G9545), Flag (Sigma®, F1804), NS1 (CTAD, Icahn School of Medicine at Mount Sinai), STAT1a (Santa Cruz, c-111), and RIG-I (Generously provided by Dr. Adolfo Garcia-Sastre, Icahn School of Medicine at Mount Sinai, NY) were used at a dilution of 1:1000 in 5% milk. Russian spring-summer encephalitis virus ascites fluid (ATCC- V558701562) was used for the detection of Langat virus at a dilution of 1:500 in 5% milk. Antibodies specific to GFP (Santa Cruz, sc-8334) and RRV-specific ascites fluid (ATCC-VR-1246AF) were used at a dilution of 1:200 in 5% milk. The SINV nsP4 antibody was a kind gift from Dr. Richard W. Hardy (Indiana University, Bloomington, IN) and was used at a 1:5,000 dilution in 5% milk in 25 mM Tris-HCl [pH 7.4], 137 mM NaCl, 2.7 mM KCl. and 0.1% Tween20. Mouse and rabbit HRP-conjugated secondary antibodies (GE Healthcare®) were used at a 1:2000 dilution in 5% milk. For antibody detection, Immobilon Western Chemoluminescent HRP (Millipore®) and SuperSignal™ West Femto Maximum Sensitivity (Thermo Fisher®) substrates were used according to the manufacture’s instructions.

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Manufacturer protocol

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