Anti-Met (c-Met) antibody [EPR22436-24]

• Flow Cyt

Unknown

Flow Cytometry - Anti-Met (c-Met) antibody [EPR22436-24] (AB216330)

Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast, Left) / B16-F10 (Mouse melanoma mixture of spindle-shaped and epithelial-like cells, Right) labeling Met (c-Met) with ab216330 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody, Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/2000 dilution.

Negative control : NIH/3T3(PMID : 21496277, 8197126, 9888438).

Gated on viable cells.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Met (c-Met) antibody [EPR22436-24] (AB216330)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized B16-F10 (mouse melanoma mixture of spindle-shaped and epithelial-like cells) and NIH/3T3 (mouse embryonic fibroblast) cells labelling Met (c-Met) with ab216330 with ab216330 at 1/50 dilution, followed by a AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining in B16-F10 cell line. Tubulin was counterstained using an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution. The nuclear counterstain was DAPI (Blue).

Negative control : NIH/3T3 (PMID : 21496277, 8197126, 9888438)

• IP

Unknown

Immunoprecipitation - Anti-Met (c-Met) antibody [EPR22436-24] (AB216330)

Met (c-Met) was immunoprecipitated from 0.35mg of B16-F10 (Mouse melanoma mixture of spindle-shaped and epithelial-like cells) whole cell lysate using ab216330 at 1/30 dilution. WB was performed on the immunoprecipitate using ab216330 at 1/1000 dilution, followed by the VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution.

Lane 1 : B16-F10 whole cell lysate 10μg (input).
Lane 2 : ab216330 IP in B16-F10 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab216330 in B16-F10 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-Met (c-Met) antibody [EPR22436-24] (ab216330)

Predicted band size: 155 kDa

false

• WB

Unknown

Western blot - Anti-Met (c-Met) antibody [EPR22436-24] (AB216330)

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

Exposure time : 26 seconds.

Negative control : NIH/3T3 (PMID : 21496277; PMID : 8197126; PMID 9888438)

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

All lanes:

Western blot - Anti-Met (c-Met) antibody [EPR22436-24] (ab216330) at 1/1000 dilution

Lane 1:

B16-F10 (mouse melanoma mixture of spindle-shaped and epithelial-like cells) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole 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

Predicted band size: 155 kDa

Observed band size: 175 kDa

false