Stat3 Antibody (F-2): sc-8019

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:200) -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 (1:200) -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

Liver allograft samples were rapidly ground in liquid nitrogen, and the cultured HSCs were harvested. The tissue powder or the isolated HSC cells were reconstituted in ice-cold RIPA buffer with protease and phosphatase inhibitors. Supernatants were recovered after centrifugation and assayed for protein content using the BCA method (Beyotime Biotechnology, P0010). Protein samples of 50 mg per lane were separated by 8% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene-difluoride (PVDF) membranes. The membranes were incubated in 5% skimmed milk for 2 h at 37°C and overnight at 4°C with primary antibodies (Santa Cruz) (rat anti-CXCR3, 1 : 200 dilution; EGFR, 1 : 200 dilution; JAK2, 1 : 200 dilution; STAT3, 1 : 200 dilution; Collagen IV, 1 : 200 dilution). GAPDH protein expression was used as a loading control. The membranes were exposed to the negative films to develop target bands after incubation with enhanced chemiluminescence (Santa Cruz, USA). The intensities of bands were quantitated by LabWorks 4.5 software (UVP, USA).

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Manufacturer protocol

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