Western blot abreview for Anti-ErbB4 / HER4 (phospho Y1284) antibody [EPR2273(2)]

Western blot

Human Cell lysate - whole cell (293 cells transfected with ErbB2-EGFP)

Reduced Denaturing (Tricine-SDS-PAGE, 7% gel)

50 µg

0.9% serum starved ~20 h, then stimulated with 20 ng/ml EGF for 5 min

293 cells transfected with ErbB2-EGFP

Milk as blocking agent for 50 minute(s) · Concentration: 5% · Temperature: ~22°C

2 hour(s) and 30 minute(s) · Diluent: 5% milk in TBS-0.1% Tween 20

1/2000

1/10000

Non-Abcam antibody was used: Donkey anti-Rabbit IgG-IRDye 800CW
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW

Fluorescence

5 minute(s) and 0 second(s)

Specific: ~200 kDa Non-specific: ~15, 20, 37, 150 kDa

No.

This experiment was to test the cross reactivity with p-ErbB2. The conclusion is that ab109273 strongly cross-reacts with p-ErbB2. There are two potential cross-reaction sites in ErbB2 (p-Y1196, p-Y1248). ErbB2-EGFP was also detected by additional ErbB2 antibodies to confirm the expression and size (not included in the figure). There is also possibility that ab109273 cross-reacts with p-EGFR (Y1138). Lane 1: EGFP transfected, 0.9% serum starved ~20 h Lane 2: ErbB2-EGFP transfected, 0.9% serum starved ~20 h Lane 3: EGFP transfected, 0.9% serum starved ~20 h, then stimulated with 20 ng EGF for 5 min Lane 4: ErbB2-EGFP transfected, 0.9% serum starved ~20 h, then stimulated with 20 ng EGF for 5 min