Anti-BIN1 抗体 [EPR13463]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BIN1 antibody [EPR13463] (AB182562)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BIN1 with purified ab182562 at 1 : 250 dilution (1 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)

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Flow Cytometry (Intracellular) - Anti-BIN1 antibody [EPR13463] (AB182562)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BIN1 with purified ab182562 at 1/30 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). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BIN1 antibody [EPR13463] (AB182562)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue sections labeling BIN1 with purified ab182562 at 1/500 dilution (0.52 μg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IP

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Immunoprecipitation - Anti-BIN1 antibody [EPR13463] (AB182562)

ab182562 (purified) at 1/20 dilution (1 µg) immunoprecipitating BIN1 in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+) : ab182562 & HeLa whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab182562 in HeLa whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-BIN1 antibody [EPR13463] (ab182562)

Predicted band size: 65 kDa

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• WB

Lab

Western blot - Anti-BIN1 antibody [EPR13463] (AB182562)

Exposure times : Lane 1 : 180 seconds; Lane 2 : 20 seconds; Lane 3 : 10 seconds.

Blocking/Dilution buffer : 5%NFDM/TBST.

All lanes:

Western blot - Anti-BIN1 antibody [EPR13463] (ab182562) at 1/5000 dilution

Lane 1:

A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 3:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 65 kDa

false

• WB

Lab

Western blot - Anti-BIN1 antibody [EPR13463] (AB182562)

The multiple bands are isoforms produced by alternative splicing

All lanes:

Western blot - Anti-BIN1 antibody [EPR13463] (ab182562) at 1/10000 dilution

Lane 1:

Human brain lysates at 15 µg

Lane 2:

Human muscle lysates at 15 µg

Secondary

All lanes:

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

Predicted band size: 65 kDa

Observed band size: 50-65 kDa

false