Flow cytometric analysis of CD18 expression on mouse splenocytes. Splenic leucocytes from C57BL/6 mice were stained with either PerCP-Cy™5.5 Rat anti-Mouse CD18 antibody (Cat. No. 562827, solid line histogram) or PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; dashed line histogram). Flow cytometric histograms were derived from gated events with the forward and side light-scattering characteristics of viable leukocytes. Flow cytometry was performed on a BD™ LSR II Flow Cytometry System.
Flow cytometric analysis of CD18 expression on mouse splenocytes. Splenic leucocytes from C57BL/6 mice were stained with either PerCP-Cy™5.5 Rat anti-Mouse CD18 antibody (Cat. No. 562827, solid line histogram) or PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; dashed line histogram). Flow cytometric histograms were derived from gated events with the forward and side light-scattering characteristics of viable leukocytes. Flow cytometry was performed on a BD™ LSR II Flow Cytometry System.
Product Details
BD Pharmingen™
Itgb2; Integrin beta-2; ITB2; LAD; LCAMB; LFA-1/Mac-1/p150,95 subunit beta
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
Cell membrane glycoproteins from mouse T-cell lymphoma BW5147
Flow cytometry (Routinely Tested)
0.2 mg/ml
16414
AB_2737822
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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- 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.
- 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–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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
550765
562827 Rev. 1
Antibody Details
C71/16
The C71/16 monoclonal antibody specifically binds to mouse CD18 which is also known as the common β2 chain of LFA-1 (CD11a/CD18, αLβ2 integrin), Mac-1 (CD11b/CD18, αMβ2 integrin), and gp150, 95 (CD11c/CD18, αxβ2 integrin). CD18 is a type I transmembrane glycoprotein. Expression of CD18 is limited to leukocytes, where it is widely distributed in consort with the three integrin α chains (CD11a, CD11b, and CD11c). Among splenocytes, NK cells have the highest density of CD18, and T lymphocytes express a higher density than the remaining cells. The β2 integrins are important mediators of leukocyte-endothelium interactions.
562827 Rev. 1
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
562827 Rev.1
Citations & References
Development References (3)
-
Springer TA. Adhesion receptors of the immune system. Nature. 1990; 346(6283):425-434. (Biology). View Reference
-
Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994; 76(2):301-314. (Biology). View Reference
-
Trowbridge IS, Omary MB. Molecular complexity of leukocyte surface glycoproteins related to the macrophage differentiation antigen Mac-1. J Exp Med. 1981; 154(5):1517-1524. (Immunogen: Flow cytometry, Immunoprecipitation). View Reference
562827 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.
