Anti-BMP4 抗体 [EPR6211]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-BMP4 antibody [EPR6211] (AB124715)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BMP4 with purified ab124715 at 1/120 dilution (10 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. 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-BMP4 antibody [EPR6211] (AB124715)

Unpurified ab124715 at 1/100 staining BMP4 in paraffin embedded Human colon tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BMP4 antibody [EPR6211] (AB124715)

Immunocytochemistry/Immunofluorescence analysis of A549 (human lung carcinoma) cells labelling BMP4 with purified ab124715 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077 Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

Secondary Only Control : PBS was used instead of the primary antibody as the negative control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BMP4 antibody [EPR6211] (AB124715)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling BMP4 with Purified ab124715 at 1 : 400 dilution (2.44 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)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-BMP4 antibody [EPR6211] (AB124715)

ab124715 (purified) at 1 : 60 dilution (2μg) immunoprecipitating BMP4 in A549 whole cell lysate.
Lane 1 (input) : A549 (Human lung carcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab124715 & A549 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab124715 in A549 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-BMP4 antibody [EPR6211] (ab124715)

Predicted band size: 47 kDa

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• WB

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Western blot - Anti-BMP4 antibody [EPR6211] (AB124715)

All lanes:

Western blot - Anti-BMP4 antibody [EPR6211] (ab124715) at 1/10000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

A549 lysate at 10 µg

Lane 3:

HeLa lysate at 10 µg

Lane 4:

A431 lysate at 10 µg

Lane 5:

Human heart lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 47 kDa

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• WB

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Western blot - Anti-BMP4 antibody [EPR6211] (AB124715)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-BMP4 antibody [EPR6211] (ab124715) at 0.05 µg/mL

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates 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: 47 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-BMP4 antibody [EPR6211] (AB124715)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• WB

CiteAb

Western blot - Anti-BMP4 antibody [EPR6211] (AB124715)

BMP4 western blot using anti-BMP4 antibody [EPR6211] ab124715. Publication image and figure legend from Li, X., Sun, B., et al., 2020, J Cancer, PubMed 32201526.

ab124715 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab124715 please see the product overview.

(A) The expression of BMP4 in four HCC cell types was evaluated according to western blot analysis. (B, C) BMP4 protein level was up-regulated in BMP4-overexpressing HepG2 cells; the up-regulation efficiency was validated through western blot and immunofluorescence. BMP4 was knocked down in SMMC7721 cells using shRNA interference; the down-regulation efficiency was also validated through western blot and immunofluorescence. Green : target protein; blue : DAPI-stained nuclear DNA. *p<0.05, scale bar=50μm.

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