Anti-BCMA antibody [EPR22457-260]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BCMA antibody [EPR22457-260] (AB253242)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U288B1 (human multiple myeloma B lymphocyte cell line) or HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells labeling BCMA with ab253242 at 1/50 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic and weak membranous staining in U266B1 cell line (PMID : 8527407) is observed.
Negative control : HEK-293T cells（PMID : 18025285).
The nuclear conter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

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

• Flow Cyt

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Flow Cytometry - Anti-BCMA antibody [EPR22457-260] (AB253242)

Flow cytometric analysis of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (left) or U266B1 (human multiple myeloma B lymphocyte cell line) (right) cells labeling BCMA with ab253242 at 1/60 dilution (red) compared with Rabbit 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.

Gated on viable cells.

Negative control : HEK-293T cells（PMID : 18025285.

• IP

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Immunoprecipitation - Anti-BCMA antibody [EPR22457-260] (AB253242)

BCMA was immunoprecipitated from 0.35 mg of U266B1 (human multiple myeloma B lymphocyte cell line) whole cell lysate with ab253242 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab253242 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

Lane 1 : U266B1 whole cell lysate 10 μg (Input).
Lane 2 : ab253242 IP in U266B1 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab253242 in U266B1 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-BCMA antibody [EPR22457-260] (ab253242)

Predicted band size: 20 kDa

Observed band size: 19 kDa,23 kDa

false

• WB

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Western blot - Anti-BCMA antibody [EPR22457-260] (AB253242)

Blocking and dilution buffer : 5% NFDM/TBST.

BCMA is a glycoprotein.
The molecular weight observed is consistent with what has been described in the literature (PMID : 23776238, 10908663,
8527407)

Negative control : HEK-293T (PMID : 18025285).

All lanes:

Western blot - Anti-BCMA antibody [EPR22457-260] (ab253242) at 1/1000 dilution

Lane 1:

U266B1 (human multiple myeloma B lymphocyte cell line) whole cell lysate at 20 µg

Lane 2:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 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: 20 kDa

Observed band size: 19 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-BCMA antibody [EPR22457-260] (AB253242)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-BCMA antibody [EPR22457-260] (ab253242) at 1/1000 dilution

All lanes:

Recombinant his-tagged human BCMA protein (<a href='/en-us/products/unavailable/recombinant-human-bcma-protein-active-ab219697'>ab219697</a>, aa1-54) 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: 20 kDa

false