Anti-BMP2 antibody [EPR24209-61]

• Flow Cyt (Intra)

Lab

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 (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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Lab

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 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. 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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IP

Lab

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) (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 (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

false

• Flow Cyt (Intra)

Lab

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 (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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Lab

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 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. 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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• WB

Lab

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 (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 44 kDa

false