Flow cytometric analysis of CD107a expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either PerCP-Cy™5.5 Rat IgG2a, λ Isotype Control (Cat. No. 560722; dashed line histogram) or PerCP-Cy™5.5 Rat Anti-Mouse CD107b antibody (Cat. No. 564248; solid line histogram). Fluorescence histograms showing CD107b expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scattering characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD107a expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either PerCP-Cy™5.5 Rat IgG2a, λ Isotype Control (Cat. No. 560722; dashed line histogram) or PerCP-Cy™5.5 Rat Anti-Mouse CD107b antibody (Cat. No. 564248; solid line histogram). Fluorescence histograms showing CD107b expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scattering characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
Lamp2; LAMP-2; LAMP II; Lysosomal-associated membrane protein 2; LGP-B
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, λ
Glycoproteins purified from BALB/c mouse embryo 3T3 cell line
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2738701
Aqueous buffered solution containing ≤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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation and Permeabilization Solution RUO
Size
125 mL
Cat No.
554722
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
564248 Rev. 2
Antibody Details
ABL-93
The ABL-93 monoclonal antibody specifically recognizes CD107b which is also known as Lysosome-associated membrane protein 2 (LAMP2/LAMP-2), Lysosomal membrane glycoprotein type B (LGP-B), and Mac-3. CD107b is a type I lysosomal transmembrane glycoprotein that serves as a useful marker to distinguish lysosomes from other cellular organelles. Mouse CD107b consists of a ~40-kDa core protein which is heavily glycosylated to form heterogeneous mature glycoproteins of 110-120 kDa. It is principally expressed in epithelial cells and macrophages in a variety of organs in normal and Beige mutant mice.
564248 Rev. 2
Format Details
PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
564248 Rev.2
Citations & References
Development References (3)
-
Cha Y, Holland SM, August JT. The cDNA sequence of mouse LAMP-2. Evidence for two classes of lysosomal membrane glycoproteins. J Biol Chem. 1990; 265(9):5008-5013. (Biology). View Reference
-
Chen JW, Chen GL, D'Souza MP, Murphy TL, August JT. Lysosomal membrane glycoproteins: properties of LAMP-1 and LAMP-2. Biochem Soc Symp. 1986; 51:97-112. (Clone-specific: Fluorescence microscopy, Immunocytochemistry (cytospins), Immunohistochemistry, Immunoprecipitation, Radioimmunoassay). View Reference
-
Chen JW, Murphy TL, Willingham MC, Pastan I, August JT. Identification of two lysosomal membrane glycoproteins. J Cell Biol. 1985; 101(1):85-95. (Immunogen: Electron microscopy, Fluorescence microscopy, Immunocytochemistry (cytospins), Immunofluorescence, Immunoprecipitation, Radioimmunoassay). View Reference
564248 Rev. 2
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.
