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R718 Mouse Anti-Human Active β2 Integrin (CD18)

BD Horizon™ R718 Mouse Anti-Human Active β2 Integrin (CD18)

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

R718 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 ethylenediaminetetraacetic acid (EDTA) anticoagulants.    Left Panel: PBL collected with heparin were either untreated (Top Plots) or were stimulated with 200 nM phorbol myristate acetate (PMA) [37°C, 20 min; Bottom Plots]. The cells were stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plots) or BD Horizon™ R718 Mouse Anti-Human Active β2 Integrin (CD18) antibody (Cat. No. 567968/567969; Right Plots). Erythrocytes were then lysed with BD FACS Lysing™ Solution (Cat. No. 349202). 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 ethylenediaminetetraacetic acid (EDTA) anticoagulants.    Left Panel: PBL collected with heparin were either untreated (Top Plots) or were stimulated with 200 nM phorbol myristate acetate (PMA) [37°C, 20 min; Bottom Plots]. The cells were stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plots) or BD Horizon™ R718 Mouse Anti-Human Active β2 Integrin (CD18) antibody (Cat. No. 567968/567969; Right Plots). Erythrocytes were then lysed with BD FACS Lysing™ Solution (Cat. No. 349202). 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 ≤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 unreacted dye was removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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 product is provided under an Agreement between BIOTIUM and BD Biosciences. 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.
  6. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567968 Rev. 1
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+.

567968 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
567968 Rev.1
Citations & References
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View product citations for antibody "567968" on CiteAb

Development References (7)

  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
  7. Wen L, Marki A, Wang Z, et al. A humanized β2 integrin knockin mouse reveals localized intra- and extravascular neutrophil integrin activation in vivo.. Cell Rep. 2022; 39(9):110876. (Clone-specific: Activation, Flow cytometry, Fluorescence activated cell sorting, Immunofluorescence, Stimulation). View Reference
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567968 Rev. 1

 

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