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BD Horizon™ BUV395 Mouse Anti-Human Active β2 Integrin (CD18)
Clone mAb 24 (also known as 24; m24; M24; mAb24)
(RUO)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 200 nM PMA (37°C, 20 min; Bottom Plots). The cells were stained with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Plots) or BD Horizon™ BUV395 Mouse Anti-Human Active β2 Integrin (CD18) antibody (Cat. No. 567947/567948; 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™ 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.
BD Horizon™ BUV395 Mouse Anti-Human Active β2 Integrin (CD18)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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. This will ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant™ Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant™ Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant™ Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant™ Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant™ Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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.
Companion Products
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+.
Development References (6)
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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
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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
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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
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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
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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
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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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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.