Anti-Met (c-Met) antibody [EPR19554-110] - BSA and Azide free

• WB

Lab

Western blot - Anti-Met (c-Met) antibody [EPR19554-110] - BSA and Azide free (AB254474)

Anti-MET antibody [EPR19554-110] (ab254252) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab254252 was shown to bind specifically to MET. A band was observed at 80-140 kDa in wild-type HeLa cell lysates with no signal observed at this size in MET knockout cell line. To generate this image, wild-type and MET knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution. The full length MET protein is glycosylated and has multiple cleavage sites, so cleaved products can be detected between ~40-140 kDa

All lanes:

Western blot - Anti-Met (c-Met) antibody [EPR19554-110] (<a href='/en-us/products/primary-antibodies/met-c-met-antibody-epr19554-110-ab254252'>ab254252</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MET (Met (c-Met)) knockout HeLa cell lysate at 20 µg

Lane 3:

HT-29 cell lysate at 20 µg

Lane 4:

T-47D cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 80-140 kDa

false

• WB

Lab

Western blot - Anti-Met (c-Met) antibody [EPR19554-110] - BSA and Azide free (AB254474)

The molecular weight observed is consistent with what has been described in the literature (PMID 8710887).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254252).

All lanes:

Western blot - Anti-Met (c-Met) antibody [EPR19554-110] (<a href='/en-us/products/primary-antibodies/met-c-met-antibody-epr19554-110-ab254252'>ab254252</a>) at 1/1000 dilution

All lanes:

B16-F10 (mouse melanoma mixture of spindle-shaped and epithelial-like cells) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 155 kDa

Observed band size: 175 kDa

false

• WB

Lab

Western blot - Anti-Met (c-Met) antibody [EPR19554-110] - BSA and Azide free (AB254474)

c-Met is an N-linked glycosylated protein. After partial de-glycosylation, the sizes of the pro c-Met (175kDa) nd P125 c-Met (125kDa) are shifted to 160, 140kDa and 80kDa (PMID : 23060267, PMID : 18187039).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254252).

All lanes:

Western blot - Anti-Met (c-Met) antibody [EPR19554-110] (<a href='/en-us/products/primary-antibodies/met-c-met-antibody-epr19554-110-ab254252'>ab254252</a>) at 1/1000 dilution

Lane 1:

Untreated B16-F10 (mouse melanoma mixture of spindle-shaped and epithelial-like cells) whole cell lysate at 10 µg

Lane 2:

B16-F10 whole cell lysate treated with PNGase F at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 155 kDa

Observed band size: 175 kDa,175-125 kDa

false

Exposure time: 37s