Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] - BSA and Azide free (AB250997)

This data was developed using ab189891, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ENO1 + ENO2 + ENO3 with ab189891 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing mostly cytoplasmic staining on Hela cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab189891 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] - BSA and Azide free (AB250997)

This data was developed using ab189891, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) labeling ENO1 + ENO2 + ENO3 with ab189891 at 1/70 dilution (red) compared with a Rabbit IgG,monoclonal - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] - BSA and Azide free (AB250997)

This data was developed using ab189891, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling ENO1 + ENO2 + ENO3 with ab189891 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing mostly cytoplasmic staining on NIH/3T3 cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody -Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab189891 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] - BSA and Azide free (AB250997)

This data was developed using ab189891, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Human ENO1 full length recombinant protein (ab89248) contains aa1-434. Human ENO2 full length recombinant protein contains aa1-434 with a His-Tag^®. Human ENO3 full length recombinant protein (ab113127) contains aa1-434 with a His-Tag^®. Human ENO2 full length recombinant protein was made in-house.

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (<a href='/en-us/products/primary-antibodies/eno1-eno2-eno3-antibody-epr18407-ab189891'>ab189891</a>) at 1/1000 dilution

Lane 1:

Human ENO1 full length recombinant protein at 0.02 µg

Lane 2:

Human ENO2 full length recombinant protein at 0.02 µg

Lane 3:

Human ENO3 full length recombinant protein at 0.02 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: 47 kDa

Observed band size: 47 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] - BSA and Azide free (AB250997)

This data was developed using ab189891, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-2 : 5 seconds; Lane 3 : 15 seconds.

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (<a href='/en-us/products/primary-antibodies/eno1-eno2-eno3-antibody-epr18407-ab189891'>ab189891</a>) at 1/1000 dilution

Lane 1:

Human fetal heart lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Lane 3:

Human fetal spleen lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

• WB

Supplier Data

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] - BSA and Azide free (AB250997)

This data was developed using ab189891, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (<a href='/en-us/products/primary-antibodies/eno1-eno2-eno3-antibody-epr18407-ab189891'>ab189891</a>) at 1/5000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

A431 (Human epidermoid carcinoma cell line) 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: 47 kDa

Observed band size: 47 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] - BSA and Azide free (AB250997)

This data was developed using ab189891, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : Lanes 1-4 : 5 seconds; Lanes 5-7 : 1 second.

Lanes 1 - 4:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (<a href='/en-us/products/primary-antibodies/eno1-eno2-eno3-antibody-epr18407-ab189891'>ab189891</a>) at 1/1000 dilution

Lanes 5 - 7:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR18407] (<a href='/en-us/products/primary-antibodies/eno1-eno2-eno3-antibody-epr18407-ab189891'>ab189891</a>) at 1/5000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Mouse heart tissue lysate at 10 µg

Lane 3:

Rat brain tissue lysate at 10 µg

Lane 4:

Rat heart tissue lysate at 10 µg

Lane 5:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 6:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 7:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell 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-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

false