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APC Rat Anti-Mouse CD49b
APC Rat Anti-Mouse CD49b
Flow cytometric analysis of CD49b on mouse splenocytes.  Splenocytes from C57BL/6 mice were stained with a FITC Mouse anti-Mouse NK1.1 antibody (clone PK136) and either with a APC Rat IgM, κ isotype control (left panel) or with the APC Rat Anti-Mouse CD49b antibody (right panel).  Dot plots were derived from gated events based on light scattering characteristics for splenocytes.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis of CD49b on mouse splenocytes.  Splenocytes from C57BL/6 mice were stained with a FITC Mouse anti-Mouse NK1.1 antibody (clone PK136) and either with a APC Rat IgM, κ isotype control (left panel) or with the APC Rat Anti-Mouse CD49b antibody (right panel).  Dot plots were derived from gated events based on light scattering characteristics for splenocytes.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Product Details
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BD Pharmingen™
itga2; Integrin alpha-2; DX5; Pan NK cell marker; VLAA2; VLA-2 alpha chain
Mouse (QC Testing)
Rat LEW, also known as Lewis IgM, κ
Mouse (C57BL/6) NK1.1+ cells propagated with rIL-2
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_1727502
Aqueous buffered solution containing protein stabilizer 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 to APC under optimum conditions, and unconjugated antibody and free APC were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560628 Rev. 2
Antibody Details
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DX5

The rat anti-mouse CD49b monoclonal antibody (clone DX5) specifically binds to the integrin α2 chain (CD49b). CD49b is a 150 kDa transmembrane glycoprotein that non-covalently associates with CD29 (integrin β1) to form the integrin α2β1 complex known as VLA-2. The rat anti-mouse CD49b antibody (clone DX5) has been reported to identify the majority of NK cells and a small T-cell subpopulation in most mouse strains (e.g., A/J, AKR, BALB/c, C3H/HeJ, C57BL/6, C57BL/10, C57BR, C58, CBA/Ca, DBA/1, DBA/2, SJL, SWR, 129/J, but not NOD). The DX5 antibody also recognizes platelets that express high levels of CD49b. Multiparameter flow cytometric analysis has demonstrated that most lymphocytes which express NK-1.1 (NKR-P1B and NKR-P1C), as detectable by mouse anti-mouse NK-1.1 antibody (clone PK136), also express the DX5 antigen. Small DX5+ NK-1.1- and DX5- NK-1.1+ cell subsets are found, especially among the CD3-positive cell population. Some CD49b+ NK cells have been reported to gradually lose reactivity with the rat anti-mouse CD49b antibody (clone DX5) when cultured in the presence of recombinant human IL-2. The resulting DX5-negative cells have weakened cytotoxic activity when compared to the remaining DX5+ cells. This indicates that the DX5 antibody distinguishes functional subsets of NK cells. No activation or blocking activity of the rat anti-mouse antibody (clone DX5) has been observed. Staining of splenic NK cells with this antibody reportedly can be blocked by hamster anti-mouse CD49b antibody (clone HMα2).

560628 Rev. 2
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 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.
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APC
Red 627-640 nm
651 nm
660 nm
560628 Rev.2
Citations & References
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Development References (12)

  1. Arase H, Saito T, Phillips JH, Lanier LL. Cutting edge: the mouse NK cell-associated antigen recognized by DX5 monoclonal antibody is CD49b (alpha 2 integrin, very late antigen-2). J Immunol. 2001; 167(3):1141-1144. (Biology: Blocking, Cytotoxicity, Flow cytometry). View Reference
  2. Chen H, Paul WE. A population of CD62Llow Nk1.1- CD4+ T cells that resembles NK1.1+ CD4+ T cells. Eur J Immunol. 1998; 28(10):3172-3182. (Biology). View Reference
  3. Gadue P, Stein PL. NK T cell precursors exhibit differential cytokine regulation and require Itk for efficient maturation. J Immunol. 2002; 169(5):2397-2406. (Biology). View Reference
  4. Hemler ME. VLA proteins in the integrin family: structures, functions, and their role on leukocytes. Annu Rev Immunol. 1990; 8:365-400. (Biology). View Reference
  5. Karnbach C, Daws MR, Niemi EC, Nakamura MC. Immune rejection of a large sarcoma following cyclophosphamide and IL-12 treatment requires both NK and NK T cells and is associated with the induction of a novel NK T cell population. J Immunol. 2001; 167(5):2569-2576. (Biology). View Reference
  6. Lee IF, Qin H, Trudeau J, Dutz J, Tan R. Regulation of autoimmune diabetes by complete Freund's adjuvant is mediated by NK cells. J Immunol. 2004; 172(2):937-942. (Biology). View Reference
  7. Miyake S, Sakurai T, Okumura K, Yagita H. Identification of collagen and laminin receptor integrins on murine T lymphocytes. Eur J Immunol. 1994; 24(9):2000-2005. (Biology). View Reference
  8. Moore TA, von Freeden-Jeffry U, Murray R, Zlotnik A. Inhibition of gamma delta T cell development and early thymocyte maturation in IL-7 -/- mice. J Immunol. 1996; 157(6):2366-2373. (Biology). View Reference
  9. Noto K, Kato K, Okumura K, Yagita H. Identification and functional characterization of mouse CD29 with a mAb. Int Immunol. 1995; 7(5):835-842. (Biology). View Reference
  10. Ortaldo JR, Winkler-Pickett R, Mason AT, Mason LH. The Ly-49 family: regulation of cytotoxicity and cytokine production in murine CD3+ cells. J Immunol. 1998; 160(1):1158-1165. (Biology). View Reference
  11. Sepulveda H, Cerwenka A, Morgan T, Dutton RW. CD28, IL-2-independent costimulatory pathways for CD8 T lymphocyte activation. J Immunol. 1999; 163(3):1133-1142. (Biology). View Reference
  12. Tanaka T, Ohtsuka Y, Yagita H, Shiratori Y, Omata M, Okumura K. Involvement of alpha 1 and alpha 4 integrins in gut mucosal injury of graft-versus-host disease. Int Immunol. 1995; 7(8):1183-1189. (Biology). View Reference
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560628 Rev. 2

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