Anti-ENPP2/ATX antibody [EPR27348-30] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ENPP2/ATX antibody [EPR27348-30] - BSA and Azide free (AB314316)

This data was developed using ab314315, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A375 (human malignant melanoma epithelial cell) cells labelling ENPP2/ATX with ab314315 at 1/50 (10.56 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in A375 cells and weak cytoplasmic staining in MCF7 cells. Low expression : MCF7 (PMID : 22140473). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-ENPP2/ATX antibody [EPR27348-30] - BSA and Azide free (AB314316)

This data was developed using ab314315, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell, Left) / A375 (human malignant melanoma epithelial cell, Right) cells labelling ENPP2/ATX with ab314315 at 1/500 dilution (0.1 ug, 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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : MCF7 (PMID : 22140473).

• WB

Supplier Data

Western blot - Anti-ENPP2/ATX antibody [EPR27348-30] - BSA and Azide free (AB314316)

This data was developed using ab314315, the same antibody clone in a different buffer formulation.

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

Low expression : skeletal muscle (PMID : 8579579).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ENPP2/ATX antibody [EPR27348-30] (<a href='/en-us/products/primary-antibodies/enpp2-atx-antibody-epr27348-30-ab314315'>ab314315</a>) at 1/1000 dilution

Lane 1:

Human ovary cancer tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 100 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-ENPP2/ATX antibody [EPR27348-30] - BSA and Azide free (AB314316)

This data was developed using ab314315, the same antibody clone in a different buffer formulation.

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

Low expression : thymus, testis (PMID : 8579579);

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1-2 : 48 seconds Lane 3-5 : 180 seconds

All lanes:

Western blot - Anti-ENPP2/ATX antibody [EPR27348-30] (<a href='/en-us/products/primary-antibodies/enpp2-atx-antibody-epr27348-30-ab314315'>ab314315</a>) at 1/1000 dilution

Lane 1:

Untreated MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 treated with 300ng/ml BFA for 24 hour whole cell lysate at 20 µg

Lane 3:

Human placenta tissue lysate at 20 µg

Lane 4:

Human thymus tissue lysate at 20 µg

Lane 5:

Human testis tissue 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

Observed band size: 100 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-ENPP2/ATX antibody [EPR27348-30] - BSA and Azide free (AB314316)

This data was developed using ab314315, the same antibody clone in a different buffer formulation.

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

Low expression : skeletal muscle (PMID : 8579579); MCF7 (PMID : 22140473).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ENPP2/ATX antibody [EPR27348-30] (<a href='/en-us/products/primary-antibodies/enpp2-atx-antibody-epr27348-30-ab314315'>ab314315</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Mouse placenta tissue lysate at 20 µg

Lane 6:

Mouse ovary tissue lysate at 20 µg

Lane 7:

Mouse skeletal muscle tissue 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

Observed band size: 100 kDa,36 kDa

false

Exposure time: 180s