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BV421 Mouse Anti-Human CD107A
BV421 Mouse Anti-Human CD107A
Flow cytometric analysis of CD107a expression on Jurkat cells (left panel).  Jurkat cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723) and subsequently stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD107a antibody (Cat. No. 562623; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact Jurkat cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Immunofluorescent analysis of CD107A expression by human cells (right panel). Cultured cells from the human U-2 OS cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050), and blocked with 5% goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in PBS. Cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD107A antibody (Cat. No. 562623, pseudo-colored green) and Alexa Fluor® 555 Mouse Anti-Human Ki-67 antibody (Cat. No. 558617, pseudo-colored red). DRAQ5 was used as a nuclear counterstain (Cat. No. 564902/564903, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 40x.
Flow cytometric analysis of CD107a expression on Jurkat cells (left panel).  Jurkat cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723) and subsequently stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD107a antibody (Cat. No. 562623; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact Jurkat cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Immunofluorescent analysis of CD107A expression by human cells (right panel). Cultured cells from the human U-2 OS cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050), and blocked with 5% goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in PBS. Cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD107A antibody (Cat. No. 562623, pseudo-colored green) and Alexa Fluor® 555 Mouse Anti-Human Ki-67 antibody (Cat. No. 558617, pseudo-colored red). DRAQ5 was used as a nuclear counterstain (Cat. No. 564902/564903, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 40x.
Product Details
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BD Horizon™
LAMP1; LAMP-1; LAMPA; LGP120
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Adult Adherent Peripheral Blood Cells
Intracellular staining (flow cytometry) (Routinely Tested), Immunofluorescence (Tested During Development)
5 µl
V P008
3916
AB_2737685
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  9. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  10. Triton is a trademark of the Dow Chemical Company.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562623 Rev. 2
Antibody Details
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H4A3

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.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

562623 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
562623 Rev.2
Citations & References
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View product citations for antibody "562623" on CiteAb

Development References (9)

  1. Alto NM, Soderling J, Scott JD. Rab32 is an A-kinase anchoring protein and participates in mitochondrial dynamics. J Biol Chem. 2002; 158(4):659-668. (Biology). View Reference
  2. Febbraio M, Silverstein RL. Identification and characterization of LAMP-1 as an activation-dependent platelet surface glycoprotein. J Biol Chem. 1990; 265(30):18531-18537. (Biology). View Reference
  3. 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
  4. Fukuda M. Lysosomal membrane glycoproteins. Structure, biosynthesis, and intracellular trafficking. J Biol Chem. 1991; 266(32):21327-21330. (Biology). View Reference
  5. 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
  6. 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
  7. 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
  8. 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.
  9. 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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562623 Rev. 2

 

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