Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12]

• Flow Cyt

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Flow Cytometry - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (AB269948)

Flow cytometric analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling Junctional Adhesion Molecule 1/JAM-A with ab269948 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 was used as the secondary antibody.

Gated on viable cells.

• Flow Cyt

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Flow Cytometry - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (AB269948)

Flow cytometric analysis of human B lymphocytes (Left) / human monocytes (Right) cells labeling Junctional Adhesion Molecule 1/JAM-A with ab269948 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 was used as the secondary antibody.

Human peripheral blood mononuclear cell (PBMC) co-stained with anti-CD19 conjugated to PE-Cy7 and anti-CD14 conjugated to BV510. JAM-A expression on B lymphocytes (CD19+) and monocytes (CD14+) population are shown respectively.

Gated on viable cells.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (AB269948)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-29 (human colorectal adenocarcinoma epithelial cell) cells labeling Junctional Adhesion Molecule 1/JAM-A with ab269948 at 1/50 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in HT-29 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - 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 : ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (AB269948)

Junctional Adhesion Molecule 1/JAM-A was immunoprecipitated from 0.35 mg human lung lysate 10μg with ab269948 at 1/30 dilution (2μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab269948 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Lane 1 : Human lung lysate 10μg.

Lane 2 : ab269948 IP in human lung lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab269948 in human lung lysate.

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

Exposure time : 15 seconds.

All lanes:

Immunoprecipitation - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

Predicted band size: 32 kDa

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• IP

Unknown

Immunoprecipitation - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (AB269948)

Junctional Adhesion Molecule 1/JAM-A was immunoprecipitated from 0.35 mg HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10μg with ab269948 at 1/30 dilution (2μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab269948 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Lane 1 : HT-29 whole cell lysate 10μg.

Lane 2 : ab269948 IP in HT-29 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab269948 in HT-29 whole cell lysate.

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

Exposure time : 15 seconds.

All lanes:

Immunoprecipitation - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

Predicted band size: 32 kDa

false

• WB

Lab

Western blot - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (AB269948)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 10698320, 10753840).

Low expression control : THP-1 and Molt-4 (PMID : 10698320).

Exposure time : Lanes 1-5 : 15 seconds; Lane 6 : 70 seconds.

All lanes:

Western blot - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948) at 1/1000 dilution

Lane 1:

Human lung lysate at 20 µg

Lane 2:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 4:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 5:

MOLT-4 (human lymphoblastic leukemia t lymphoblast) whole cell lysate at 20 µg

Lane 6:

TF-1 (human erythroleukemia erythroblast) 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 1/50000 dilution

Predicted band size: 32 kDa

Observed band size: 35 kDa

false