Flow cytometric analysis of CD107b 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), permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723), and subsequently stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Human CD107b antibody (Cat. No. 565306; solid line histogram). The fluorescence histogram showing CD107b expression (or Ig Isotype control staining) was derived from events with the forward and side light-scatter characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD LSRII Cell Analyzer System.
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Flow cytometric analysis of CD107b 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), permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723), and subsequently stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Human CD107b antibody (Cat. No. 565306; solid line histogram). The fluorescence histogram showing CD107b expression (or Ig Isotype control staining) was derived from events with the forward and side light-scatter characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD LSRII Cell Analyzer System.
Product Details
BD Pharmingen™
LAMP-2; LAMP2; LAMPB; LGP110
Human (QC Testing)
Mouse IgG1, κ
Human Adult Adherent Peripheral Blood Cells
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
V P007
AB_2739172
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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
PE Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554680
565306 Rev. 1
Antibody Details
H4B4
The H4B4 monoclonal antibody specifically binds to CD107b. CD107b is a heavily glycosylated, ~120 kDa type I transmembrane protein that is also known as lysosomal-associated membrane protein 2 (LAMP-2/LAMP2), LAMPB, or LGP110. CD107b is expressed in the lysosomal and endosomal membranes of most cell types where it may play a role in maintaining membrane integrity. CD107b is also expressed on the surface of activated platelets and lymphocytes, including T lymphocytes, e.g., degranulating CD8+ cytotoxic T cells. CD107b serves as a ligand for selectins, including E-selectin, and mediates cellular adhesion. CD107a/ LAMP-1 and CD107b /LAMP-2 are carriers for poly-N-acetyllactosamines, and display the sialyl Le[x] antigen. CD107b is likewise expressed on some tumor cells and cell lines, including U937 and KG1a, and may facilitate tumor metastases.
565306 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
565306 Rev.1
Citations & References
Development References (9)
-
Azorsa DO. Hildreth, JEK. CD107a (LAMP-1) and CD107b (LAMP-2) cluster workshop report. In: 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; 1995:1351.
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Chan KS, Kaur A. Flow cytometric detection of degranulation reveals phenotypic heterogeneity of degranulating CMV-specific CD8+ T lymphocytes in rhesus macaques. J Immunol Methods. 2007; 325(1-2):20-34. (Clone-specific: Flow cytometry). View Reference
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Chen JW, Cha Y, Yuksel KU, Gracy RW, August JT. Isolation and sequencing of a cDNA clone encoding lysosomal membrane glycoprotein mouse LAMP-1. Sequence similarity to proteins bearing onco-differentiation antigens. J Biol Chem. 1988; 263(18):8754-8758. (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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Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
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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.
565306 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
