ErbB2 (HER-2) Antibody Cocktail

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	-Mouse (1 µg/mL)

Protocol tips
-Mouse (1 µg/mL)

Publication protocol

Protein extracts were separated using SDS-PAGE under denaturing conditions (4–20% Mini-PROTEAN TGX Gels) and were transferred to nitrocellulose membranes (Bio-Rad Laboratories). Membranes were blocked with 5% milk-TBST (Tris-buffered saline and 0.1% Tween 20) and incubated with the indicated primary antibodies overnight at +4 °C. Primary antibodies were diluted in blocking buffer (Thermo, StartingBlock (PBS) blocking, #37538) and PBS (1:1 ratio) mix and incubated overnight at +4oC. Primary antibody dilutions used ranged from 1:500 to 1:1000. After primary antibody incubation, membranes were washed three times with TBST and incubated with fluorophore-conjugated secondary antibodies diluted (1:1000) in blocking buffer at RT for 1 h. Membranes were scanned using an infrared imaging system (Odyssey; LI-COR Biosciences). The following secondary antibodies were used: donkey anti-mouse IRDye 800CW (LI-COR, 926-32212), donkey anti-mouse IRDye 680RD (LI-COR, 926-68072), donkey anti-rabbit IRDye 800CW (LI-COR, 926-32213) and donkey anti-rabbit IRDye 680RD (LI-COR, 926-68073).

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Manufacturer protocol

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