Anti-BIN1 antibody [EPR13463]

6 product images

• 

  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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab182562 in HeLa whole cell lysate
  For western blotting, VeriBlot for IP Detection Reagent (HRP) (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

• 

  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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 65 kDa

• 

  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 (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.

• 

  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

• 

  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, Goat Anti-Rabbit IgG H&L (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).

• 

  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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (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.