Anti-BMP2 antibody [EPR24209-61]

6 product images

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  Immunoprecipitation - Anti-BMP2 antibody [EPR24209-61] (ab284387)

  BMP2 was immunoprecipitated from 0.35 mg Saos-2 (human osteosarcoma epithelial) whole cell lysate 10 ug with ab284387 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab284387 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Saos-2 (human osteosarcoma epithelial) whole cell lysate 10 ug

  Lane 2: ab284387 IP in Saos-2 whole cell lysate

  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab284387 in Saos-2 whole cell lysate

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

  Exposure time: 24 seconds.

  All lanes: Immunoprecipitation - Anti-BMP2 antibody [EPR24209-61] (ab284387)

  Predicted band size: 44 kDa

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  Western blot - Anti-BMP2 antibody [EPR24209-61] (ab284387)

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

  The molecular weight observed is consistent with literature.

  Bands observed at 14KD are monomers (PMID: 12819188, PMID: 3200834).

  Bands observed at 18KD are glycosylated monomers (PMID: 12819188, PMID: 8117284, PMID: 16261447, PMID: 3200834).

  Bands observed at ~30KD are dimers or glycosylated dimers) (PMID: 22508088, PMID: 16261447, PMID: 3200834).

  Bands observed at ~38kD is another isoform (PMID: 20230640)

  Exposure time: Lanes 1-2: 37 seconds; Lanes 3-6: 8 seconds; Lanes 7-8: 37 seconds

  All lanes: Western blot - Anti-BMP2 antibody [EPR24209-61] (ab284387) at 1/1000 dilution

  Lane 1: Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: Saos-2 (human osteosarcoma epithelial) whole cell lysate at 20 µg

  Lane 3: LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

  Lane 5: RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

  Lane 6: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

  Lane 7: Mouse lung tissue lysate at 20 µg

  Lane 8: Rat spleen tissue lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

  Predicted band size: 44 kDa

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  Flow Cytometry (Intracellular) - Anti-BMP2 antibody [EPR24209-61] (ab284387)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling BMP2 with ab284387 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

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  Flow Cytometry (Intracellular) - Anti-BMP2 antibody [EPR24209-61] (ab284387)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Caco-2 (human colorectal adenocarcinoma epithelial cell) cells labelling BMP2 with ab284387 at 1/500 dilution (0.1ug)/ (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

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  Immunocytochemistry/ Immunofluorescence - Anti-BMP2 antibody [EPR24209-61] (ab284387)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling BMP2 with ab284387 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cells is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

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  Immunocytochemistry/ Immunofluorescence - Anti-BMP2 antibody [EPR24209-61] (ab284387)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Caco-2 cells labelling BMP2 with ab284387 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Caco-2 cells is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.