Recombinant Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23244-12] to Junctional Adhesion Molecule 1/JAM-A
- Suitable for: ICC/IF, WB, Flow Cyt, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12]
See all Junctional Adhesion Molecule 1/JAM-A primary antibodies -
Description
Rabbit monoclonal [EPR23244-12] to Junctional Adhesion Molecule 1/JAM-A -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, Flow Cyt, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human lung tissue lysate. HT-29, HUVEC and TF-1 whole cell lysate. ICC/IF: HT-29 cells. Flow Cyt: HT-29 cells, human B lymphocytes and human monocytes. IP: HT-29 whole cell lysate. Human lung tissue lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23244-12 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab269948 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC/IF |
1/50.
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WB |
1/1000. Predicted molecular weight: 32 kDa.
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Flow Cyt |
1/500.
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IP |
1/30.
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Notes |
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ICC/IF
1/50. |
WB
1/1000. Predicted molecular weight: 32 kDa. |
Flow Cyt
1/500. |
IP
1/30. |
Target
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Function
Seems to plays a role in epithelial tight junction formation. Appears early in primordial forms of cell junctions and recruits PARD3. The association of the PARD6-PARD3 complex may prevent the interaction of PARD3 with JAM1, thereby preventing tight junction assembly (By similarity). Plays a role in regulating monocyte transmigration involved in integrity of epithelial barrier. Involved in platelet activation. In case of orthoreovirus infection, serves as receptor for the virus. -
Sequence similarities
Belongs to the immunoglobulin superfamily.
Contains 2 Ig-like V-type (immunoglobulin-like) domains. -
Post-translational
modificationsN-glycosylated. -
Cellular localization
Cell junction > tight junction. Cell membrane. Localized at tight junctions of both epithelial and endothelial cells. - Information by UniProt
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Database links
- Entrez Gene: 50848 Human
- Omim: 605721 Human
- SwissProt: Q9Y624 Human
- Unigene: 517293 Human
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Alternative names
- CD 321 antibody
- CD321 antibody
- CD321 antigen antibody
see all
Images
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All lanes : Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948) at 1/1000 dilution
Lane 1 : Human lung lysate
Lane 2 : HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 4 : HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 5 : MOLT-4 (human lymphoblastic leukemia t lymphoblast) whole cell lysate
Lane 6 : TF-1 (human erythroleukemia erythroblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 32 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?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.
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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.
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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.
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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.
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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.
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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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab269948 has been referenced in 1 publication.
- Deng L et al. Bioengineered miR-124-3p prodrug selectively alters the proteome of human carcinoma cells to control multiple cellular components and lung metastasis in vivo. Acta Pharm Sin B 11:3950-3965 (2021). PubMed: 35024318