Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (AB180943)

Immunocytochemistry/ Immunofluorescence analysis of Ramos (human Burkitt's lymphoma B lymphocyte) cells labeling Tcl1 with purified ab225718 at 1/50 (2.6 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-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 (blue) was used as 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-ENO1 + ENO2 + ENO3 antibody [EPR12483] (AB180943)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed U87-MG cells labeling NSE with ab180943 at 1/10 dilution (red) compared to a Rabbit monoclonal IgG Isotype control (green), followed by Goat anti rabbit IgG (FITC) secondary antibody at 1/150 dilution.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (AB180943)

Immunocytochemistry/ Immunofluorescence analysis of Raji (human Burkitt's lymphoma B lymphocyte) cells labeling CD23 with purified ab135386 at 1/25 (7.48 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IP

Supplier Data

Immunoprecipitation - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (AB180943)

Western blot analysis of HepG2 cell lysate immunoprecipitated using ab180943 at 1/50 dilution (Lane 1). Lane 2 : Negative control. Anti-Rabbit IgG (HRP) secondary antibody, specific to the non-reduced form of IgG, used at 1/1500 dilution.

All lanes:

Immunoprecipitation - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (ab180943)

Predicted band size: 47 kDa

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• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (AB180943)

Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/100(1.65 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

Lab

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (AB180943)

Western blot : Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] ab180943 staining at 1/1000 dilution, shown in green; Mouse anti-6x HisTag (ab18184) loading control staining at 1/20,000 dilution, shown in magenta.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution. For DDDDK-tag detection, HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (ab49763) was used at 1/1000 dilution and developed using the ECL technique.

Exposure time for DDDDK tag : 0.1s.

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (ab180943) at 1/1000 dilution

Lane 1:

ENO1 Recombinant Protein (His Tag) at 0.2 µg

Lane 2:

ENO2 Recombinant Protein (His Tag) at 0.2 µg

Lane 3:

ENO3 Recombinant Protein (His Tag) at 0.2 µg

Lane 4:

ENO1 Recombinant Protein (DDDDK Tag) at 0.2 µg

Lane 5:

ENO2 Recombinant Protein (DDDDK Tag) at 0.2 µg

Lane 6:

ENO3 Recombinant Protein (DDDDK Tag) at 0.2 µg

Secondary

Lanes 1 - 6:

Goat anti-Mouse 680RD at 1/20000 dilution

Lanes 1 - 6:

Goat anti-Rabbit 800CW at 1/20000 dilution

Observed band size: 50 kDa

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• WB

Supplier Data

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (AB180943)

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (ab180943) at 1/5000 dilution

Lane 1:

Fetal brain lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Lane 3:

SH-SY5Y cell lysate at 20 µg

Lane 4:

U87-MG cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 47 kDa

Observed band size: 47 kDa

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