CD107a (LAMP-1) Monoclonal Antibody (eBioH4A3), PE-Cyanine5, eBioscience™

Product Specific Information

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 predominantly 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. It has also been reported for use in surface staining in a flow cytometric based degranulation assay.

Applications Tested: This eBioH4A3 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of normal normal human peripheral blood cells. 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.

Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488-561 nm; Emission: 667 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

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.