Two-color flow cytometric analysis of CD9 expression on mouse bone marrow cells. Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD11b antibody (Cat. No. 553311/557397/561689) and either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Panel) or Alexa Fluor® 647 Rat Anti-Mouse CD9 antibody (Cat. No. 564233; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD9 (or Ig Isotype control staining) versus CD11b were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-color flow cytometric analysis of CD9 expression on mouse bone marrow cells. Mouse bone marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD11b antibody (Cat. No. 553311/557397/561689) and either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Left Panel) or Alexa Fluor® 647 Rat Anti-Mouse CD9 antibody (Cat. No. 564233; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD9 (or Ig Isotype control staining) versus CD11b were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
Cd9; Tetraspanin-29; Tspan29
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
(C57BL/6 x DBA/2)F1 mouse bone marrow-derived stromal cell line BMS2
Flow cytometry (Routinely Tested)
0.2 mg/ml
12527
AB_2738687
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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
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
Alexa Fluor® 647 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557690
PE Rat Anti-CD11b RUO
Size
0.2 mg
Cat No.
553311
564233 Rev. 1
Antibody Details
KMC8
The KMC8 monoclonal antibody specifically binds to CD9 which is also known as Tetraspanin-29 (Tspan29). CD9 is a 24-kDa member of the transmembrane 4 superfamily, also called the tretraspanin family. In the mouse, CD9 is present on bone marrow myeloid cells, stromal cells, and megakaryocyte-committed progenitors; subsets of peripheral T and B lymphocytes; and neutrophils, platelets, dendritic cells, and bone marrow-derived macrophages. CD9 has been found to be associated with integrins and other cell-surface receptors. It appears to play roles in signal transduction and in regulating cellular adhesive properties. CD9 can also reportedly participate in T-cell costimulation and induction of apoptosis. The KMC8 antibody reportedly blocks certain CD9 functions and activates macrophages.
564233 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
564233 Rev.1
Citations & References
Development References (9)
-
Jennings LK, Crossno JT Jr, Fox CF, White MM, Green CA. Platelet p24/CD9, a member of the tetraspanin family of proteins. Ann N Y Acad Sci. 1994; 714:175-184. (Biology). View Reference
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Kaji K, Takeshita S, Miyake K, Takai T, Kudo A. Functional association of CD9 with the Fc gamma receptors in macrophages. J Immunol. 2001; 166(5):3256-3265. (Clone-specific: Activation). View Reference
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Miyake K, Medina KL, Hayashi S, Ono S, Hamaoka T, Kincade PW. Monoclonal antibodies to Pgp-1/CD44 block lympho-hemopoiesis in long-term bone marrow cultures. J Exp Med. 1990; 171(2):477-488. (Immunogen: ELISA). View Reference
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Nakorn TN, Miyamoto T, Weissman IL. Characterization of mouse clonogenic megakaryocyte progenitors. Proc Natl Acad Sci U S A. 2003; 100(1):205-210. (Biology). View Reference
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Oritani K, Wu X, Medina K, et al. Antibody ligation of CD9 modifies production of myeloid cells in long-term cultures. Blood. 1996; 87(6):2252-2261. (Immunogen: Blocking). View Reference
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Park CS, Yashiro Y, Tai XG, et al. Differential involvement of a Fas-CPP32-like protease pathway in apoptosis of TCR/CD9-costimulated, naive T cells and TCR-restimulated, activated T cells.. J Immunol. 1998; 160(12):5790-6. (Clone-specific: Apoptosis, (Co)-stimulation). View Reference
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Pulendran B, Lingappa J, Kennedy MK, et al. Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice. J Immunol. 1997; 159(5):2222-2231. (Biology). View Reference
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Tanio Y, Yamazaki H, Kunisada T, Miyake K, Hayashi SI. CD9 molecule expressed on stromal cells is involved in osteoclastogenesis. Exp Hematol. 1999; 27(5):853-859. (Clone-specific: Blocking). View Reference
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Wright MD, Tomlinson MG. The ins and outs of the transmembrane 4 superfamily. Immunol Today. 1994; 15(12):588-594. (Biology). View Reference
564233 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.
