JAM-B/VE-JAM Antibody [Unconjugated]

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.

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.

JAM‑B/VE‑JAM was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human/Mouse JAM‑B/VE‑JAM Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1074) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane in endothelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

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.

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.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, IHC, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

JAM-B/VE-JAM Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human JAM‑B/VE-JAM
  Phe29-Asn236
  Accession # P57087
• Specificity: Detects human JAM‑B/VE-JAM in direct ELISAs. Detects human and mouse JAM-B/VE-JAM in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: JAM2
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-25 ug/mL
  • Neutralization 0.2-0.8 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 1-2 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for JAM-B/VE-JAM Antibody [Unconjugated]

• 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

Background

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.

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for JAM-B/VE-JAM Antibody (AF1074)(2)

Publications using AF1074

• 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 Junctional adhesion molecule (JAM)-B supports lymphocyte rolling and adhesion through interaction with alpha4beta1 integrin. Immunology, 2009-10-01;128(2):196-205. 2009-10-01 [PMID: 19740376] (ICC, Human)
• Nagasawa K, Chiba H, Fujita H, Kojima T, Saito T, Endo T, Sawada N Possible involvement of gap junctions in the barrier function of tight junctions of brain and lung endothelial cells. J. Cell. Physiol., 2006-07-01;208(1):123-32. 2006-07-01 [PMID: 16547974] (ICC, Western Blot, Porcine)

Review for JAM-B/VE-JAM Antibody (AF1074) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer Flow Human 01/05/2015

Summary

• Application: Flow Cytometry
• Sample Tested: See PMID 19795504
• Species: Human

Bioinformatics

• Gene Symbol: JAM2
• Uniprot:
  • P57087()