Anti-ENO1 antibody [EPR19758]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 antibody [EPR19758] (AB227978)

Immunohistochemical analysis of paraffin-embedded human clear cell renal cell carcinoma tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and nuclear staining in human clear cell kidney carcinoma (PMID : 26037892) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ENO1 antibody [EPR19758] (AB227978)

Immunofluorescent analysis of 4% paraformaldegyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling ENO1 with ab227978 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining in HeLa cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

-ve control : PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution .

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 antibody [EPR19758] (AB227978)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of human pancreas (PMID : 19425054) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ENO1 antibody [EPR19758] (AB227978)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling ENO1 with ab227978 at 1/600 dilution (red) compared witha Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).

Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-ENO1 antibody [EPR19758] (AB227978)

ENO1 was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab227978 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227978 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Jurkat whole cell lysate 10 μg (Input).

Lane 2 : ab227978 IP in Jurkat whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab227978 in Jurkat whole cell lysate.

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-ENO1 antibody [EPR19758] (ab227978)

Predicted band size: 47 kDa

Observed band size: 47 kDa

true

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ENO1 antibody [EPR19758] (AB227978)

Immunofluorescent analysis of 4% paraformaldegyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling ENO1 with ab227978 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in NIH/3T3 cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

-ve control : PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluo^r® 488) (ab150077) secondary antibody at 1/1000 dilution .

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 antibody [EPR19758] (AB227978)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of mouse pancreas is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENO1 antibody [EPR19758] (AB227978)

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling ENO1 with ab227978 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in medium-small ducts of rat pancreas is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

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Western blot - Anti-ENO1 antibody [EPR19758] (AB227978)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lane 1 : 15 seconds.

Lanes 2-7 : 1 second.

All lanes:

Western blot - Anti-ENO1 antibody [EPR19758] (ab227978) at 1/1000 dilution

Lane 1:

Human skeletal muscle tissue lysate at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

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

Lane 5:

C6 (rat glial tumor glial 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 embryo 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 1/100000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

true

• WB

Unknown

Western blot - Anti-ENO1 antibody [EPR19758] (AB227978)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ENO1 antibody [EPR19758] (ab227978) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Mouse heart tissue lysate at 10 µg

Lane 3:

Mouse kidney tissue lysate at 10 µg

Lane 4:

Rat brain tissue lysate at 10 µg

Lane 5:

Rat heart tissue lysate at 10 µg

Lane 6:

Rat liver tissue lysate at 10 µg

Lane 7:

Rat spleen tissue 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 1/100000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

true

Exposure time: 3min