Anti-BMP2 antibody [EPR20807]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BMP2 antibody [EPR20807] (AB214821)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized LNCaP (human prostate cancer cell line) cells labeling BMP2 with ab214821 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on LNCaP cell line.

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

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-BMP2 antibody [EPR20807] (AB214821)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line labeling BMP2 with ab214821 at 1/700 dilution (red) compared with an Isotype controlRabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black)and an unlabelled 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BMP2 antibody [EPR20807] (AB214821)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling BMP2 with ab214821 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on RAW 264.7 cell line.

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

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

• WB

Supplier Data

Western blot - Anti-BMP2 antibody [EPR20807] (AB214821)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-4 : 3 minutes; Lane 5 : 30 seconds; Lane 6 : 5 seconds.

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).

All lanes:

Western blot - Anti-BMP2 antibody [EPR20807] (ab214821) at 1/1000 dilution

Lane 1:

LNCaP (human prostate cancer cell line) whole cell lysate at 20 µg

Lane 2:

Mouse cartilage tissue lysate at 20 µg

Lane 3:

Human lung cancer tissue lysate at 20 µg

Lane 4:

Human fetal kidney tissue lysate at 20 µg

Lane 5:

Rat spleen tissue lysate at 10 µg

Lane 6:

C6 (rat glial tumor 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 1/20000 dilution

Predicted band size: 44 kDa

Observed band size: 14 kDa,18 kDa,30 kDa

true

• WB

Supplier Data

Western blot - Anti-BMP2 antibody [EPR20807] (AB214821)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 3 minutes; Lanes 2-4 : 15 seconds.

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).

All lanes:

Western blot - Anti-BMP2 antibody [EPR20807] (ab214821) at 1/1000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 2:

Saos-2 (human osteosarcoma cell line) whole cell lysate at 10 µg

Lane 3:

C2C12 (mouse myoblast cell line) whole cell lysate at 10 µg

Lane 4:

RAW264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) 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 1/20000 dilution

Predicted band size: 44 kDa

Observed band size: 14 kDa,18 kDa,30 kDa

true

• WB

Supplier Data

Western blot - Anti-BMP2 antibody [EPR20807] (AB214821)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-BMP2 antibody [EPR20807] (ab214821) at 1/1000 dilution

Lane 1:

His-tagged human BMP2 (aa283-395) recombinant protein at 0.02 µg

Lane 2:

His-tagged human BMP4 (aa293-408) recombinant protein at 0.02 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: 44 kDa

Observed band size: 15 kDa

true

Exposure time: 1s