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PE-CF594 Rat Anti-Mouse CD49b
PE-CF594 Rat Anti-Mouse CD49b
Multicolor flow cytometric analysis of CD49b expressed on mouse splenocytes. Splenocytes from C57BL/6 mice were stained with a FITC Mouse Anti-Mouse NK1.1 antibody (Cat. No. 553857/561067) and with either BD Horizon™ PE-CF594 Rat IgM, κ Isotype Control (Cat. No. 562489, Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse CD49b antibody (Cat. No. 562453, Right Panel). Two color flow cytometric dot plots showing the expression of NK1.1 versus CD49b (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of CD49b expressed on mouse splenocytes. Splenocytes from C57BL/6 mice were stained with a FITC Mouse Anti-Mouse NK1.1 antibody (Cat. No. 553857/561067) and with either BD Horizon™ PE-CF594 Rat IgM, κ Isotype Control (Cat. No. 562489, Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse CD49b antibody (Cat. No. 562453, Right Panel). Two color flow cytometric dot plots showing the expression of NK1.1 versus CD49b (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
製品詳細
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BD Horizon™
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_11153857
Aqueous buffered solution containing BSA, 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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. CF™ is a trademark of Biotium, Inc.
  10. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  11. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  12. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
562453 Rev. 1
抗体の詳細
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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).

This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red (eg 610/20-nm filter).

562453 Rev. 1
フォーマットの詳細
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, 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 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-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.
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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
562453 Rev.1
引用&参考文献
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Development References (3)

  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. (Clone-specific: Blocking, Cytotoxicity, Flow cytometry). View Reference
  2. 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
  3. 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
562453 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.