Anti-Met (c-Met) antibody [EPR19067]

• WB

Lab

Western blot - Anti-Met (c-Met) antibody [EPR19067] (AB216574)

For Western blotting of c-Met, we recommend ab227637 instead of this product.

ab227637 detects both the full-length c-Met and the β-chain, including β-chain degradation products between 35-55 kDa, and generally produces cleaner Western blot results than ab216574.

c-Met can undergo glycosylation, resulting in a higher molecular weight. It can also be cleaved into α and β chains (PMID : 34210960).

The β-chain undergoes degradation, resulting in lower molecular weight fragments (PMID : 32522525).

Blocking and diluting buffer and concentration : 5% NFDM /TBST.

Lanes 1 - 2:

Western blot - Anti-Met (c-Met) antibody [SP44] - C-terminal (<a href='/ko/products/primary-antibodies/met-c-met-antibody-sp44-c-terminal-ab227637'>ab227637</a>) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution

Lanes 1 and 3:

Wild-type HeLa cell lysate at 20 µg

Lanes 2 and 4:

MET knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35-55 kDa,145-175 kDa

false

Exposure time: 60s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Met (c-Met) antibody [EPR19067] (AB216574)

ELISA analysis of Human c-met recombinant protein at 1000 ng/mL with ab216574. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-Met (c-Met) antibody [EPR19067] (AB216574)

Anti-Met (c-Met) antibody [EPR19067] (ab216574) staining at 1/1000 dilution, shown in green; Mouse GAPDH (ab8245) loading control staining at 1/20000 dilution, shown in magenta. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution. Please note that we offer an alternative anti-c-MET antibody, ab227637, which has shown lower background and cleaner Western blot performance in our internal studies.

All lanes:

Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-met-met-c-met-knockout-hela-cell-line-ab265961'>ab265961</a>) 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 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 155 kDa,121 kDa

Observed band size: 182 kDa,139 kDa

false

• WB

Lab

Western blot - Anti-Met (c-Met) antibody [EPR19067] (AB216574)

Lanes 1 - 2 : Merged signal (red and green). Green - ab216574 observed at 156 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab216574 was shown to react with Met (c-Met) in wild-type HeLa. Loss of signal was observed when knockout cell line ab265961 (knockout cell lysate ab256991) was used. Wild-type and Met (c-Met) knockout samples were subjected to SDS-PAGE. ab216574 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution

All lanes:

Western blot - Human MET (Met (c-Met)) knockout HeLa cell pellet (<a href='/ko/products/cell-lysates/human-met-met-c-met-knockout-hela-cell-pellet-ab278899'>ab278899</a>)

Predicted band size: 155 kDa

Observed band size: 156 kDa

false

• WB

Supplier Data

Western blot - Anti-Met (c-Met) antibody [EPR19067] (AB216574)

Blocking/Dilution buffer : 5% NFDM/TBST.

In human liver the antibody detected c-Met beta subunit (145 kDa) [PMID : 22418436] and a cleavage c-Met fragment (100 kDa) [PMID : 18187039].

All lanes:

Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution

All lanes:

Human liver lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 155 kDa

Observed band size: 100-150 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Met (c-Met) antibody [EPR19067] (AB216574)

Blocking and Diluting buffer and concentration : 5% NFDM /TBST

All lanes:

Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/10000 dilution

Lane 1:

293T whole cell lysate (Human epithelial cell line from embryonic kidney) transfected with an empty expression vector at 10 µg

Lane 2:

293T whole cell lysate transfected with a His-tagged human c-Met construct at 10 µg

Secondary

All lanes:

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

Predicted band size: 155 kDa

Observed band size: 150-175 kDa

false

Exposure time: 1s