Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free (AB220216)

This data was developed using ab79757, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling ENO1 + ENO2 + ENO3 with purified ab79757 at 1/500 dilution (0.20 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free (AB220216)

This data was developed using ab79757, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of U-87 MG (Human glioblastoma-astrocytoma epithelial cell) cells labeling ENO1 + ENO2 + ENO3 with purified ab79757 at 1/50 dilution (2.0 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free (AB220216)

This data was developed using ab79757, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling NSE with purified ab79757 at 1/20 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free (AB220216)

This data was developed using ab79757, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling ENO1 + ENO2 + ENO3 with purified ab79757 at 1/500 dilution (0.20 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free (AB220216)

This data was developed using ab79757, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling ENO1 + ENO2 + ENO3 with purified ab79757 at 1/500 dilution (0.20 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Lab

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free (AB220216)

This data was developed using ab79757, the same antibody clone in a different buffer formulation.

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

Anti-DDDDK tag antibody [6F7] - Loading Control (ab205606) staining at 1/1000 dilution.

The cross-reactivity with ENO2 is weaker than that with ENO1 and ENO3

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/eno1-eno2-eno3-antibody-epr3377-neuronal-marker-ab79757'>ab79757</a>) at 1/1000 dilution

Lane 1:

Recombinant human ENO1 protein with C-DDDDK tag at 10 ng

Lane 2:

Recombinant human ENO2 protein with C-DDDDK tag at 10 ng

Lane 3:

Recombinant human ENO3 protein with C-DDDDK tag at 10 ng

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: 50 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free (AB220216)

This data was developed using ab79757, the same antibody clone in a different buffer formulation.

Mouse anti-6x HisTag was used as a loading control.

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/eno1-eno2-eno3-antibody-epr3377-neuronal-marker-ab79757'>ab79757</a>) at 1/1000 dilution

Lane 1:

Human ENO3 protein (His Tag) at 0.2 µg

Lane 2:

Human ENO2 protein (His Tag) at 0.2 µg

Lane 3:

Human ENO1 protein (His Tag) at 0.2 µg

Predicted band size: 47 kDa

false

• WB

Unknown

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - BSA and Azide free (AB220216)

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - Neuronal Marker (<a href='/en-us/products/primary-antibodies/eno1-eno2-eno3-antibody-epr3377-neuronal-marker-ab79757'>ab79757</a>) at 1/1000 dilution

Lane 1:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Y79 (Human retinoblastoma retinoblastoma) whole cell lysate at 20 µg

Lane 4:

Mouse brain lysate at 20 µg

Lane 5:

Rat brain 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

Predicted band size: 47 kDa

false