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PE-CF594 Mouse Anti-Human Active β2 Integrin (CD18)
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BD Horizon™ PE-CF594 Mouse Anti-Human Active β2 Integrin (CD18)

Clone mAb 24 (also known as 24; m24; M24; mAb24)

(RUO)
PE-CF594 Mouse Anti-Human Active β2 Integrin (CD18)
Multiparameter flow cytometric analysis of Active β2 Integrin (CD18) expression on untreated or stimulated human peripheral blood leucocytes (PBL) collected with either heparin or EDTA anticoagulants.      Left Panel: PBL collected with heparin were either untreated (Top Plots) or were stimulated with 125 ng/ml Phorbol 12-Myristate 13-Acetate (PMA; 37°C, 20 min; Bottom Plots). The cells were stained with either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; Left Plots) or BD Horizon™ PE-CF594 Mouse Anti-Human Active β2 Integrin (CD18) antibody (Cat. No. 567956/567957; Right Plots). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plots showing the correlated expression of Active β2 Integrin (CD18) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.      Right Panel: PBL collected with the chelating agent, EDTA were similarly treated, stained, and analyzed by multiparameter flow cytometry.
Multiparameter flow cytometric analysis of Active β2 Integrin (CD18) expression on untreated or stimulated human peripheral blood leucocytes (PBL) collected with either heparin or EDTA anticoagulants.      Left Panel: PBL collected with heparin were either untreated (Top Plots) or were stimulated with 125 ng/ml Phorbol 12-Myristate 13-Acetate (PMA; 37°C, 20 min; Bottom Plots). The cells were stained with either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; Left Plots) or BD Horizon™ PE-CF594 Mouse Anti-Human Active β2 Integrin (CD18) antibody (Cat. No. 567956/567957; Right Plots). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plots showing the correlated expression of Active β2 Integrin (CD18) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.      Right Panel: PBL collected with the chelating agent, EDTA were similarly treated, stained, and analyzed by multiparameter flow cytometry.
Product Details
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BD Horizon™
ITGB2; Integrin beta-2; Integrin β2; LFA-1 β; LFA1b; beta 2 integrin; β2 integrin
Human (QC Testing)
Mouse BALB/c IgG1, κ
Fibronectin-purified Human Monocytes
Flow cytometry (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA 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 the dye under optimum conditions and unconjugated antibody and free dye were removed.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  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.
  13. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Antibody Details
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mAb 24

mAb 24 (also known as 24 or m24) specifically recognizes a divalent cation-dependent epitope expressed by active human Integrin β2 which is also known as CD18. Integrin β2 is an ~95 kDa type I transmembrane glycoprotein that is encoded by ITGB2 (integrin subunit beta 2). Integrin β2 (CD18) noncovalently associates with other integrin α chains to form αLβ2 (CD11a/CD18; also known as, LFA-1), αMβ2 (CD11b/CD18; Mac-1, CR3), αXβ2 (CD11c/CD18; p150,95; CR4) and αDβ2 (CD11d/CD18) heterodimers. These integrins are variably expressed on lymphocytes, NK cells, neutrophils, eosinophils, basophils, monocytes, macrophages, Langerhans cells, and dendritic cells (DCs). These leucocyte integrins bind to various ligands and mediate cellular adhesion and signaling responses that regulate cellular activation, effector function, and migration. mAb 24 binds to the extended/open, high-affinity ligand-binding conformation of active Integrin β2 (CD18). mAb 24 can be used as a reporter antibody for the activated status of Integrin β2-containing receptors expressed by leucocytes in response to various stimuli in the presence of Mg2+ or Mn2+.

Format Details
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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
Citations & References
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View product citations for antibody "567957" on CiteAb

Development References (6)

  1. Chen X, Xie C, Nishida N, Li Z, Walz T, Springer TA. Requirement of open headpiece conformation for activation of leukocyte integrin alphaXbeta2.. Proc Natl Acad Sci U S A. 2010; 107(33):14727-32. (Clone-specific). View Reference
  2. Dransfield I, Cabañas C, Craig A, Hogg N. Divalent cation regulation of the function of the leukocyte integrin LFA-1.. J Cell Biol. 1992; 116(1):219-26. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
  3. Dransfield I, Hogg N. Regulated expression of Mg2+ binding epitope on leukocyte integrin alpha subunits.. EMBO J. 1989; 8(12):3759-65. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
  4. Hogg N, Selvendran Y. An anti-human monocyte/macrophage monoclonal antibody, reacting most strongly with macrophages in lymphoid tissue.. Cell Immunol. 1985; 92(2):247-53. (Immunogen: Flow cytometry, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Radioimmunoassay). View Reference
  5. Kamata T, Tieu KK, Tarui T, Puzon-McLaughlin W, Hogg N, Takada Y. The role of the CPNKEKEC sequence in the beta(2) subunit I domain in regulation of integrin alpha(L)beta(2) (LFA-1).. J Immunol. 2002; 168(5):2296-301. (Clone-specific: Flow cytometry). View Reference
  6. Lefort CT, Rossaint J, Moser M, et al. Distinct roles for talin-1 and kindlin-3 in LFA-1 extension and affinity regulation.. Blood. 2012; 119(18):4275-82. (Clone-specific: Flow cytometry). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.