Anti-COL11A1 antibody [EPR24327-8]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-COL11A1 antibody [EPR24327-8] (AB270946)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A-204 (Human Muscle Rhabdomyosarcoma cell line) whole cells labeling COL11A1 with ab270946 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining in A-204 cell line. Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab195889) at 1/200 dilution (Red).

Negative control : HCT 116 (PMID : 25417197).

The nuclear counterstain is DAPI (blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-COL11A1 antibody [EPR24327-8] (AB270946)

Intracellular flow cytometric analysis of HCT 116 (Human colorectal carcinoma epithelial cell), Left)/ A-204 (rat muscle rhabdomyosarcoma, Right) fixed with 4% paraformaldehyde permeabilized with 90% methanol cells labelling COL11A1 with ab270946 at 1/500 (Red), compared with a (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 was used as the secondary antibody.

Negative control : HCT116 (PMID : 25417197).

• WB

Lab

Western blot - Anti-COL11A1 antibody [EPR24327-8] (AB270946)

Blocking and diluting buffer : 5% NFDM/TBST.

Lysates were prepared from fresh material and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-COL11A1 antibody [EPR24327-8] (ab270946) at 1/1000 dilution

Lane 1:

A-204 (Human muscle rhabdomyosarcoma) transfected with scrambled siRNA control, whole cell fresh lysate at 20 µg

Lane 2:

A-204 transfected with siRNA specifically targeti COL11A1, whole cell fresh lysate at 20 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 1/10000 dilution

Predicted band size: 181 kDa

Observed band size: 260 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-COL11A1 antibody [EPR24327-8] (AB270946)

Blocking and dilution buffer : 5% NFDM/TBST.

Lysates were prepared from fresh material and used for Western blotting immediately to minimize protein degradation.

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 32950601).

The ~150 kDa bands are expected to be non-specific bands.

Negative control : HCT116 (PMID : 25417197).

All lanes:

Western blot - Anti-COL11A1 antibody [EPR24327-8] (ab270946) at 1/1000 dilution

Lane 1:

A-204 (Human muscle rhabdomyosarcoma) whole cell fresh lysate at 20 µg

Lane 2:

HCT116 (human colorectal carcinoma epithelial cell) whole cell fresh lysate at 20 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 1/20000 dilution

Predicted band size: 181 kDa

Observed band size: 280 kDa

true

Exposure time: 3min