Anti-DGAT1 antibody [EPR13430-4] - N-terminal

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling DGAT1 with Purified ab181180 at 1 : 100 dilution (4.3 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling DGAT1 with Purified ab181180 at 1/400 dilution (1μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

Supplier Data

Western blot - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

All lanes:

Western blot - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (ab181180) at 1/50000 dilution

All lanes:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

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• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

Immunofluorescence analysis of acetone-fixed HeLa cells, staining DGAT1 (green) with ab181180 at 1/100 dilution. Alexa Fluor®488-conjugated goat anti rabbit IgG was used as a secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

Intracellular Flow Cytometry analysis of DGAT1 expression in HeLa cells using ab181180 at 1/70 dilution (red) and a rabbit IgG as negative control (green).

• IP

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Immunoprecipitation - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

ab181180 (purified) at 1 : 30 dilution (2μg) immunoprecipitating DGAT1 in Human fetal kidney lysate.
Lane 1 (input) : Human fetal kidney lysate 10μg
Lane 2 (+) : ab181180 & Human fetal kidney lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab181180 in Human fetal kidney lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (ab181180)

Predicted band size: 55 kDa

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• IP

Supplier Data

Immunoprecipitation - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

Western blot analysis on immunoprecipitation pellet from Human fetal kidney lysate (lane 1) or negative control (lane 2), labeling DGAT1 immunoprecipitated using ab181180 at 1/1500 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

All lanes:

Immunoprecipitation - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (ab181180)

Predicted band size: 55 kDa

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

Blocking buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (ab181180) at 1/10000 dilution

All lanes:

Human small intestine tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

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Exposure time: 3min

• WB

Lab

Western blot - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

Blocking buffer and concentration : 5% NFDM/TBST

We recommend to use 1% SDS Hot lysis method to get clear Western Blot results.

All lanes:

Western blot - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (ab181180) at 1/10000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate prepared using RIPA lysis method at 20 µg

Lane 2:

HEK-293 whole cell lysate prepared using 1% SDS Hot lysis method at 20 µg

Lane 3:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate prepared using RIPA lysis method at 20 µg

Lane 4:

Jurkat whole cell lysate prepared using 1% SDS Hot lysis method at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (AB181180)

All lanes:

Western blot - Anti-DGAT1 antibody [EPR13430-4] - N-terminal (ab181180) at 1/10000 dilution

All lanes:

Human fetal kidney 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-ab136636'>ab136636</a>) at 1/500 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

false