CD107a (LAMP-1) Monoclonal Antibody (eBioH4A3), eFluor™ 450, eBioscience™, Invitrogen™

Description

Description: The eBioH4A3 monoclonal antibody reacts with human CD107a, also known as lysosomal-associated membrane protein-1 (LAMP-1). CD107a is a highly glycosylated protein of approximately 110kDa. It is predomitly expressed intracellularly in the lysosomal/endosomal membrane in nearly all cells. CD107a is transiently expressed on the cell surface of degranulating cytolytic T cells, and is also upregulated on the surface of activated platelets and some cancer cells. Applications Reported: This eBioH4A3 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This eBioH4A3 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of Jurkat cells using the Intracellular Fixation & Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at 5 μL (0.25 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. eFluor™ 450 is an alternative to Pacific Blue™. eFluor™ 450 emits at 445 nm and is excited with the Violet laser (405 nm).

LAMP1 (CD107a, lysosome-associated membrane protein-1) together with LAMP-2, is a major constituent of lysosomal membrane, 1-2% of total CD107a is found also on the plasma membrane. LAMP1 is a heavily glycosylated membrane protein which contains a putative signal peptide, 18 sites for N-linked glycosylation, a single membrane-spanning segment and a short (11 amino acid) cytosolic tail. The LAMP proteins are involved in lysosome biogenesis and are required for fusion of lysosomes with phagosomes. LAMP1 is a type 1 integral membrane protein that is transported from trans-Golgi network to endosomes and then lysosomes. Upon cell activation, LAMP1 transfer to the plasma membrane is dependent on a carboyxl-terminal tyrosine based motif (YXXI). Perturbation in the spacing between the tyrosine based motif relative to the membrane abolishes lysosome localization of LAMP1, and this mutant protein then cycles between the plasma membrane and the endosome. Cell surface LAMP1 (and LAMP2) have been shown to promote adhesion of human peripheral blood mononuclear cells (PBMC) to vascular endothelium, therefore, they are possibly involved in the adhesion of PBMC to the site of inflammation. Increased LAMP1 immunoreactivity is observed in neurons and glial cells surrounding senile plaques in Alzheimer’s Disease (AD) cases, and is localized in medullary epithelial cells, single macrophages and lymphocytes in acute thymic involution. LAMP1 is a good marker of mast cell activation.