Ub Antibody (P4D1): sc-8017

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

Samples solubilized in 1× Laemmli sample buffer, containing 5% β-mercaptoethanol were incubated at 37°C for 30 min, separated in 4%–20% MOPS SDS-PAGE gels, and transferred to PVDF membranes using the Bio-Rad system. PVDF membranes were first blocked in TBS-T buffer (50 mM Tris-HCl, 150 mM NaCl, and 0.2% Tween 20) containing 5% dry milk for 2 h and then incubated with the indicated primary antibody in the blocking buffer overnight, washed three times for 10 min each, and incubated with horseradish peroxidase-conjugated secondary antibodies (either goat anti-mouse, goat anti-rabbit, or goat anti-rat 1:5,000–10,000). Following all antibody incubations, membranes were washed three times in TBS-T buffer, and protein bands were visualized using the Prime Western blotting detection system (GE HealthCare). Primary antibodies include rabbit anti-TH (EMD-Millipore AB152, 1:1,000), rabbit anti-DAT “EL2” (EMD-Millipore AB5802, 1:1,000), rat anti-DAT “MAB” (EMD-Millipore MAB369, 1:1,000), mouse anti-SUMO1 (Santa Cruz sc-5308, 1:1,000), mouse anti-HA antibody (Santa Cruz sc-7392, 1:1,000), mouse anti-ubiquitin (Santa Cruz sc-8017, 1:1,000), rabbit anti-Ubc9 (Santa Cruz sc-10759, 1:1,000), and mouse anti- β-actin (EMD-Millipore MAB1501, 1:5,000). The PageRuler pre-stained protein ladder was used to estimate protein molecular weights on immunoblots (Thermo 26616).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Western blotting Ubiquitin using Ub Antibody (P4D1): sc-8017 from Santa Cruz Biotechnology.

Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for Ub Antibody (P4D1): sc-8017 below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting Ubiquitin using Ub Antibody (P4D1): sc-8017 from Santa Cruz Biotechnology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.