Smad1 Antibody (A-4): sc-7965

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

For Western blot, SO, NO, O kidneys were removed from wild type and S-ENG+ mice [WT: SO (n = 6); NO (n = 8); O (n = 8) and S-ENG+: SO (n = 6); NO (n = 8); O (n = 8)] and immediately stored at − 80 °C. Tissue protein extracts were prepared as previously described [16]. Total kidney protein lysates (60 μg) were fractioned by SDS-PAGE, transferred onto PVDF membrane (Millipore Billerica, MA, USA) and incubated overnight at 4 °C with the antibodies for endoglin, fibronectin, collagen I, α-SMA, PCNA, Smad1, Smad2/3, phospho-Smad1, phospho-Smad3, ICAM-1, VCAM-1, COX-2, GAPDH, β-actin, ALK1 and ALK5 in the conditions specified in Table 2. Development was performed using a chemiluminescent reagent (ECL detection reagent, Amersham Biosciences) and signals were recorded on X-ray film (Fujifilm), followed by densitometric analysis (Scion Image).

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