Anti-Wnt6 antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Wnt6 antibody (AB50030)

ICC/IF image of ab50030 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab50030, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• WB

Project

Western blot - Anti-Wnt6 antibody (AB50030)

Wnt6 has a predicted molecular weight of 40 kDa; however it has a number of potential glycosylation sites which may affect the migration of the protein (SwissProt data). Abcam currently sell two antibodies to this target, ab49946 which detects a band at 78 kDa and ab50030 which detects a band at 55 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-Wnt6 antibody (ab50030) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 3:

A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HEK293 Human embryonic kidney cell line Whole Cell Lysate at 10 µg

Lane 5:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 6:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 7:

SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

Lane 8:

U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 40 kDa

Observed band size: 55 kDa

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