Human Ubiquitin/Ubiquitin+1 Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Bring all the solutions and reagents to room temperature before use; otherwise the detection limit may be compromised.	-Rabbit (1:2000) -Always wear gloves or use forceps when handling blotting membranes to avoid contamination with protein from fingers.

Upstream tips
-Bring all the solutions and reagents to room temperature before use; otherwise the detection limit may be compromised.
Protocol tips
-Rabbit (1:2000) -Always wear gloves or use forceps when handling blotting membranes to avoid contamination with protein from fingers.

Publication protocol

The cells were lysed in ATM lysis buffer (containing 100 mM Tris-Cl [pH 7.5], 150 mM NaCl, 0.2 mM EDTA, 20% glycerol, 0.4% NP-40, 2% Tween-20 and 0.2 mM PMSF). The protein concentration in the supernatant was measured using a BCA Assay Kit (Novagen, #71285-3). Then, the samples were loaded into a 10% or 15% gel to resolve the proteins. Different amounts of total protein were loaded in each experiment to facilitate the detection of different target proteins. After electrophoresis, the proteins were transferred to PVDF membranes (Amersham, #10600021) and hybridized with primary antibodies at a dilution of 1:2000. The HRP-labeled secondary antibodies (Zhongshan Golden Bridge) were applied at a dilution of 1:4000. An ECL detection system (Calbiochem, #345818) was used to detect the signals on the membranes.

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Manufacturer protocol

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