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PE Mouse Anti-Human Active β2 Integrin (CD18)
PE 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 200 nM PMA (37°C, 20 min; Bottom Plots). The cells were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plots) or PE Mouse Anti-Human Active β2 Integrin (CD18) antibody (Cat. No. 567770; Right Plots). Erythrocytes were then lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). 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 viable 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.

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 PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plots) or PE Mouse Anti-Human Active β2 Integrin (CD18) antibody (Cat. No. 567770; Right Plots). Erythrocytes were then lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). 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 viable 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.

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BD Pharmingen™
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


准备和存储

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.

推荐的实验流程

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.

商品通知

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  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. An isotype control should be used at the same concentration as the antibody of interest.
  7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
抗体详情
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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+.

        

格式详情
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE 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 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
报价单和参考
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研发参考 (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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
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