Reactivity | HuSpecies Glossary |
Applications | Bioactivity |
Format | Carrier-Free |
Details of Functionality | Measured by the ability of the immobilized protein to support the adhesion of BT‑20 human breast cancer cells. When 5 x 104 cells/well are added to rhAMICA/Fc Chimera coated plates (10 µg/mL, 100 µL/well), approximately 55%-75% will adhere after 1 hour at 37° C. Similar results were obtained using HeLa, A549, and T84 cell-lines. Optimal dilutions should be determined by each laboratory for each application. |
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Source | Mouse myeloma cell line, NS0-derived human AMICA/JAML protein
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Accession # | |||||||
N-terminal Sequence | Leu10 |
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Structure / Form | Disulfide-linked homodimer |
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Protein/Peptide Type | Recombinant Proteins |
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Gene | JAML |
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Purity | >90%, by SDS-PAGE under reducing conditions and visualized by silver stain |
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Endotoxin Note | <0.10 EU per 1 μg of the protein by the LAL method. |
Dilutions |
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Theoretical MW | 55.5 kDa (monomer). Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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SDS-PAGE | 65-70 kDa, reducing conditions |
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Publications |
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Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Buffer | Lyophilized from a 0.2 μm filtered solution in PBS. |
Purity | >90%, by SDS-PAGE under reducing conditions and visualized by silver stain |
Reconstitution Instructions | Reconstitute at 100 μg/mL in sterile PBS. |
AMICA (adhesion molecule, interacting with CXADR antigen 1), also known as JAML, is a 65 kDa, heavily glycosylated transmembrane protein that belongs to the junctional adhesion molecule (JAM) subset of the immunoglobulin superfamily (1). JAM family molecules contribute to intercellular connections within epithelial and endothelial cell layers, and mediate their interactions with various hemopoietic cells (1). The human AMICA cDNA encodes a 384 amino acid (aa) precursor that includes a 19 aa signal sequence, a 256 aa extracellular domain (ECD) with two Ig-like domains, a 21 aa transmembrane segment, and a 98 aa cytoplasmic domain (2). Alternative splicing may generate isoforms with N- and C-terminal deletions. In contrast to other JAM family proteins, AMICA does not contain a cytoplasmic PDZ-binding motif (3). Within the ECD, human AMICA shares 58% and 63% aa sequence identity with mouse and rat AMICA, respectively. It shares 18% - 20% aa sequence identity with the ECDs of human JAM-A, -B, -C, and JAM4. AMICA is expressed on the surface of granulocytes and monocytes and is upregulated during the differentiation of myeloid leukemia cells (2, 3). A motif in the ECD, which promotes dimerization of other JAM family proteins, is required for surface localization of AMICA (2). AMICA mediates the adhesion of monocytes to endothelial cells (2) and neutrophil migration across epithelial cell monolayers (3). This latter function involves specific interactions of AMICA with the coxsackie virus and adenovirus receptor (CXADR) in epithelial tight junctions (3). In particular, the membrane proximal Ig-like domain of AMICA binds the membrane-distal Ig-like domain of CXADR (3). AMICA does not appear to interact homophilically, as neutrophils adhere to immobilized CXADR but not to immobilized AMICA (3).
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