Anti-LAMC2 antibody [EPR23654-127] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-LAMC2 antibody [EPR23654-127] - BSA and Azide free (AB274384)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 cells labelling LAMC2 with ab274376 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in A431 cell line. Low expression control : A549 (PMID : 25591736). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274376).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-LAMC2 antibody [EPR23654-127] - BSA and Azide free (AB274384)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized A549 (Human lung carcinoma epithelial cell) (Left) A431 (Human epidermoid carcinoma epithelial cell) (Right) cells labelling LAMC2 with ab274376 at 1/500 dilution (0.1ug) (Red) (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) at 1/2000 dilution was used as the secondary antibody.

Low expression control : A549.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274376).

• IP

Unknown

Immunoprecipitation - Anti-LAMC2 antibody [EPR23654-127] - BSA and Azide free (AB274384)

LAMC2 was immunoprecipitated from 0.35 mg A431 (human epidermoid carcinoma epithelial cell), whole cell lysate with ab274376 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab274376 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : A431 (human epidermoid carcinoma epithelial cell), whole cell lysate 10 ug

Lane 2 : ab274376 IP in A431 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab274376 in A431 whole cell lysate.

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

Exposure time : 41 seconds

Band of 140KDa represents the full length form, Bands of 100KDa and 80KDa represent the endogenously cleaved form.

The expression profile observed is consistent with what has been described in the literature (PMID : 21829200).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274376).

All lanes:

Immunoprecipitation - Anti-LAMC2 antibody [EPR23654-127] (<a href='/en-us/products/primary-antibodies/lamc2-antibody-epr23654-127-ab274376'>ab274376</a>)

Predicted band size: 131 kDa

Observed band size: 100 kDa,140 kDa,80 kDa

false

• WB

Lab

Western blot - Anti-LAMC2 antibody [EPR23654-127] - BSA and Azide free (AB274384)

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

Bands of 140KDa and 80KDa respectively represent the full length and endogenously cleaved form.

The expression profile observed is consistent with what has been described in the literature (PMID : 25591736, 21829200).

Low expression : A549 (PMID : 25591736) and WI-38 (PMID : 10964684)

Expsure time : 3 minutes

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274376).

All lanes:

Western blot - Anti-LAMC2 antibody [EPR23654-127] (<a href='/en-us/products/primary-antibodies/lamc2-antibody-epr23654-127-ab274376'>ab274376</a>) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

WI-38 (human fetal lung fibroblast), whole cell 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 1/100000 dilution

Predicted band size: 131 kDa

Observed band size: 140 kDa,80 kDa

false