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. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5.We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) 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 nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered. CD107a (lysosome-associated membrane protein-1, LAMP-1), together with LAMP-2, is a major constituent of lysosomal membrane, 1-2% of total CD107a is found also on the plasma membrane. The LAMP proteins are involved in lysosome biogenesis and are required for fusion of lysosomes with phagosomes. Increased CD107a immunoreactivity is observed in neurones, and in glial cells surrounding senile plaques in Alzheimers disease cases and is localized mainly in medullary epithelial cells, single macrophages and lymphocytes in acute thymic involution. CD107a is a good marker of mast cell activation.
|Store at 2-8°C. Do not freeze. Light-sensitive material. This tandem dye is sensitive to photo-induced oxidation. Protect this vial from light during storage|
|PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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