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BD OptiBuild™ RY775 Mouse Anti-Human CD18 (Integrin β2)
Clone mAb 24 (also known as 24; m24; M24; mAb24)
(RUO)
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 to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- 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 (7)
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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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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
Please refer to Support Documents for Quality Certificates
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.