Human/Mouse JAM-B/VE-JAM Antibody

(2 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human JAM‑B/VE-JAM in direct ELISAs. Detects human and mouse JAM-B/VE-JAM in Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human JAM‑B/VE-JAM
    Phe29-Asn236
    Accession # P57087
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1-2 µg/mL
    See below
  • Simple Western
    20 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize JAM‑B/VE‑JAM-mediated adhesion of the J45.01 human acute lymphoblastic leukemia T lymphocyte cell line. Fong, S. et al. (2002) J. Immunol. 168:1618. The Neutralization Dose (ND50) is typically 0.2-0.8 µg/mL in the presence of 0.2 µg/mL Recombinant Human JAM‑B/VE‑JAM Fc Chimera.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human JAM‑B/VE‑JAM by Western Blot. Western blot shows lysates of human placenta tissue and human testis tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse JAM‑B/VE‑JAM Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1074) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for JAM‑B/VE‑JAM at approximately 43-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Mouse JAM‑B/VE‑JAM by Western Blot. Western blot shows lysates of bEnd.3 mouse endothelioma cell line and SVEC4‑10 mouse vascular endothelial cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human/Mouse JAM‑B/VE‑JAM Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1074) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for JAM‑B/VE‑JAM at approximately 40-48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human JAM‑B/VE‑JAM by Simple WesternTM. Simple Western lane view shows lysates of human placenta tissue and human testis tissue, loaded at 0.2 mg/mL. A specific band was detected for JAM‑B/VE‑JAM at approximately 58-61 kDa (as indicated) using 20 µg/mL of Goat Anti-Human JAM‑B/VE‑JAM Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1074) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Cell Adhesion Mediated by JAM‑B/VE‑JAM and Neutral­ization by Human JAM‑B/VE‑JAM Antibody. Recombinant Human JAM‑B/VE‑JAM Fc Chimera (Catalog # 1074-VJ), immobilized onto a microplate previously coated with Goat Anti-Human IgG Fc (Catalog # G-102-C), supports the adhesion of the J45.01 human acute lymphoblastic leukemia T lymphocyte cell line in a dose-dependent manner (orange line), as measured by endogenous cellular lysosomal acid phosphatase activity. Adhesion elicited by Recombinant Human JAM‑B/VE‑JAM Fc Chimera (0.2 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Mouse JAM-B/VE-JAM Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1074). The ND50 is typically 0.2-0.8 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: JAM-B/VE-JAM

The family of juctional adhesion molecules (JAM), comprising at least three members, are type I transmembrane receptors belonging to the immunoglobulin (Ig) superfamily (1, 2). These proteins are localized in the tight junctions between endothelial cells or epithelial cells. Some family members are also found on blood leukocytes and platelets. JAM-B, alternatively named vascular endothelial JAM (VE-JAM), is expressed prominently on high endothelial venules of lymphoid organs where it is localized to the intercellular boundaries of high endothelial cells. It is also expressed on the endothelium of a variety of non-lymphoid organs, especially the heart and placenta (3, 5). Human JAM-B cDNA predicts a 298 amino acid residue (aa) precursor protein with a putative 28 aa signal peptide, a 209 aa extracellular region containing two Ig domains, a 23 aa transmembrane domain and a 38 aa cytoplasmic domain containing a PDZ-binding motif and a PKC phosphorylation site. Human JAM-B shares approximately 79% aa sequence homology with its mouse homologue. It also shares approximately 35% aa sequence homology with human JAM-A or JAM-C. JAM-B exhibits homotypic interactions, as well as heterotypic interactions with JAM-C, but not JAM-A (4, 5, 7). It is also a ligand for the integrin alpha 4 beta 1. However, the JAM-B/ alpha 4 beta 1 interaction is facilitated only after prior adhesion of JAM-B to JAM-C (6). Through its heterotypic interactions with JAM-C, JAM-B is an adhesive ligand for T, NK, and dendritic cells, and may play a role in regulating leukocyte transmigration (5).

The nomenclature used for the JAM family proteins is confusing. VE-JAM has been referred in the literature variously as JAM-B or JAM-3. Until further clarification, R&D Systems has adopted the nomenclature where both mouse and human VE-JAM are referred to as JAM-B.

  • References:
    1. Chavakis, T. et al. (2003) Thromb. Haemost. 89:13.
    2. Aurand-Lions, M. et al. (2001) Blood 98:3699.
    3. Palmeri, A. et al. (2000) J. Biol. Chem. 275:19139.
    4. Cunnigham, S. et al. (2000) J. Biol. Chem. 275:34750.
    5. Liang, T. et al. (2002) J. Immunol. 168:1618.
    6. Cunningham, A. et al. (2002) J Biol. Chem. 277:27589.
    7. Arrate, M. et al. (2001) J. Biol. Chem. 276:45826.
  • Long Name:
    Junctional Adhesion Molecule B
  • Entrez Gene IDs:
    58494 (Human); 67374 (Mouse)
  • Alternate Names:
    C21orf43; CD322 antigen; CD322; JAM2; JAMB; JAM-B; JAM-BJAM-IT/VE-JAM; junctional adhesion molecule 2JAM-2; junctional adhesion molecule B; PRO245; Vascular endothelial junction-associated molecule; VE-JAM; VE-JAMVEJAMCD322
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Junctional adhesion molecule (JAM)-B supports lymphocyte rolling and adhesion through interaction with alpha4beta1 integrin.
    Authors: Ludwig RJ, Hardt K, Hatting M, Bistrian R, Diehl S, Radeke HH, Podda M, Schon MP, Kaufmann R, Henschler R, Pfeilschifter JM, Santoso S, Boehncke WH
    Immunology, 2009;128(2):196-205.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  2. Possible involvement of gap junctions in the barrier function of tight junctions of brain and lung endothelial cells.
    Authors: Nagasawa K, Chiba H, Fujita H, Kojima T, Saito T, Endo T, Sawada N
    J. Cell. Physiol., 2006;208(1):123-32.
    Species: Porcine
    Sample Type: Cell Lysates
    Application: WB
Isotype Controls
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Normal Goat IgG Control

Ctrl AB-108-C 191  
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Secondary Antibodies
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Goat IgG HRP-conjugated Antibody

WB, Simple Western HAF109 19  
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Goat IgG HRP-conjugated Antibody

WB HAF017 15  
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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 9
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 6  
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Donkey Anti-Goat IgG Antibody

WB AF109 6
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Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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