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Flow cytometric analysis of CD107a expression in Jurkat cells. Jurkat cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723) The cells were subsequently stained with either Alexa Fluor® 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; dashed line histogram) or Alexa Fluor® 488 Mouse anti-Human CD107a antibody (Cat. No. 567006/567007; solid line histogram) in BD Perm/Wash™ Buffer. The fluorescence histogram showing CD107a expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact Jurkat cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ Alexa Fluor® 488 Mouse Anti-Human CD107a
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Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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Companion Products
The H4A3 monoclonal antibody specifically binds to CD107a which is also known as Lysosomal-associated membrane protein 1 (LAMP-1). LAMP-1 is a ~110 kDa type I transmembrane protein that is heavily glycosylated and widely expressed by cells primarily on the luminal surface of their lysosomes. It is also expressed on the surface of activated platelets, activated lymphocytes, cytotoxic T cells and NK cells, and some tumor cell lines, including U937 and KG1a. LAMP-1 can serve as a ligand for E-selectin-mediated cell adhesion. LAMP-1 and LAMP-2 (CD107b) are carriers for poly-N-acetyllactosamines and are able to display sialyl Le[x] termini.
Development References (9)
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Fukuda M, Viitala J, Matteson J, Carlsson SR. Cloning of cDNAs encoding human lysosomal membrane glycoproteins, h-lamp-1 and h-lamp-2. Comparison of their deduced amino acid sequences. J Biol Chem. 1988; 263(35):18920-18928. (Biology). View Reference
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Hocking DC, Kowalski K. A cryptic fragment from fibronectin's III1 module localizes to lipid rafts and stimulates cell growth and contractility. J Cell Biol. 2002; 158(1):175-184. (Biology). View Reference
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Mane SM, Marzella L, Bainton DF, et al. Purification and characterization of human lysosomal membrane glycoproteins. Arch Biochem Biophys. 1989`; 268(1):360-378. (Immunogen: Electron microscopy, Flow cytometry, Immunoaffinity chromatography, Immunohistochemistry, Immunoprecipitation). View Reference
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Sawada R, Lowe JB, Fukuda M. E-selectin-dependent adhesion efficiency of colonic carcinoma cells is increased by genetic manipulation of their cell surface lysosomal membrane glycoprotein-1 expression levels. J Biol Chem. 1993; 268(17):12675-12681. (Biology). View Reference
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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Spoerl Z, Stumpf M, Noegel AA, Hasse A. Oligomerization, F-actin interaction, and membrane association of the ubiquitous mammalian coronin 3 are mediated by its carboxyl terminus. J Biol Chem. 2002; 277(50):48858-48867. (Biology: Blocking). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.