Flow cytometric analysis of CD107a (LAMP-1) expression by Jurkat cells. Cells from the Human Jurkat (Acute T cell leukemia, ATCC® TIB-152™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with either BD Horizon™ RY610 Mouse IgG1, κ Isotype Control (Cat. No. 571151; dashed line histogram) or BD Horizon™ RY610 Mouse Anti-Human CD107a (LAMP-1) antibody (Cat. No. 571485/571493; solid line histogram). The fluorescence histogram showing CD107a (LAMP-1) 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™ X-20 Cell Analyzer System and FlowJo™ Software.
Flow cytometric analysis of CD107a (LAMP-1) expression by Jurkat cells. Cells from the Human Jurkat (Acute T cell leukemia, ATCC® TIB-152™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with either BD Horizon™ RY610 Mouse IgG1, κ Isotype Control (Cat. No. 571151; dashed line histogram) or BD Horizon™ RY610 Mouse Anti-Human CD107a (LAMP-1) antibody (Cat. No. 571485/571493; solid line histogram). The fluorescence histogram showing CD107a (LAMP-1) 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™ X-20 Cell Analyzer System and FlowJo™ Software.
Product Details
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)
5 µl/test
V P008
3916
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
RY610 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
571151
571485 Rev. 1
Antibody Details
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.
571485 Rev. 1
Format Details
RY610
The BD Horizon RealYellow™ 610 (RY610) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 610-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY610 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY610 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE-CF594.
RY610
Yellow-Green 561 nm
557 nm
610 nm
571485 Rev.1
Citations & References
View product citations for antibody "571485" on CiteAb
Development References (11)
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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
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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
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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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Fukuda M. Lysosomal membrane glycoproteins. Structure, biosynthesis, and intracellular trafficking. J Biol Chem. 1991; 266(32):21327-21330. (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, Fluorescence activated cell sorting, 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: Intracellular Staining/Flow Cytometry). 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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Trotta E, Bessette PH, Silveria SL, et al. A human anti-IL-2 antibody that potentiates regulatory T cells by a structure-based mechanism.. Nat Med. 2018; 24(7):1005-1014. (Clone-specific: Flow cytometry). View Reference
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Willemen Y, Van den Bergh JM, Bonte SM, et al. The tumor-associated antigen RHAMM (HMMR/CD168) is expressed by monocyte-derived dendritic cells and presented to T cells.. Oncotarget. 2016; 7(45):73960-73970. (Clone-specific: Flow cytometry). View Reference
571485 Rev. 1
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.