Multiparameter flow cytometric analysis of CD11b (Integrin αM) expression on viable Mouse bone marrow cells. BALB/c Mouse bone-marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with either BD Horizon™ RB705 Rat IgG2b, κ Isotype Control (Cat. No. 570263; Left Plot) or BD Horizon™ RB705 Rat Anti-CD11b (Integrin αM) antibody (Cat. No. 570292/570293; Right Plot) at 0.125 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD11b (Integrin αM) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD11b (Integrin αM) expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either BD Horizon™ RB705 Rat IgG2b, κ Isotype Control (Cat. No. 570263; Left Plot) or BD Horizon™ RB705 Rat Anti-CD11b (Integrin αM) antibody (Cat. No. 570292/570293; Right Plot) at 0.125 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of CD11b (Integrin αM) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD11b (Integrin αM) expression on viable Mouse bone marrow cells. BALB/c Mouse bone-marrow cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with either BD Horizon™ RB705 Rat IgG2b, κ Isotype Control (Cat. No. 570263; Left Plot) or BD Horizon™ RB705 Rat Anti-CD11b (Integrin αM) antibody (Cat. No. 570292/570293; Right Plot) at 0.125 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD11b (Integrin αM) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD11b (Integrin αM) expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either BD Horizon™ RB705 Rat IgG2b, κ Isotype Control (Cat. No. 570263; Left Plot) or BD Horizon™ RB705 Rat Anti-CD11b (Integrin αM) antibody (Cat. No. 570292/570293; Right Plot) at 0.125 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of CD11b (Integrin αM) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
製品詳細
BD Horizon™
Itgam; Integrin alpha-M; Ly-40; Mac-1a; Mac-1 alpha; CR3A; CR-3 alpha chain
Mouse (QC Testing), Human (Tested in Development)
Rat DA, also known as DA/HA IgG2b, κ
Mouse Splenic Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
3684, 16409
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation and Storage
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
推奨アッセイ手順
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
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
570292 Rev. 1
抗体の詳細
M1/70
The M1/70 monoclonal antibody specifically binds to CD11b, also known as Integrin alpha M (Itgam or αM). CD11b is a 170-kDa type 1 transmembrane glycoprotein and belongs to the Integrin alpha chain family. CD11b serves as the alpha chain of the heterodimeric Mac-1 integrin (CD11b/CD18, αMβ2), also known as complement receptor 3 (CR3). Mac-1 mediates adhesion to ICAM-1 (CD54), ICAM-2 (CD102), fibrinogen and binding to C3bi. Mac-1 is expressed at varying levels on granulocytes, macrophages, myeloid-derived dendritic cells, natural killer cells, microglia, and B-1 B lymphocytes. Mac-1 expression is rapidly upregulated on neutrophils after activation, in the same time period that CD62L (L-selectin) is shed from the cell surface. The M1/70 antibody reportedly blocks cell adherence and C3bi binding but does not block cell-mediated lysis. Cross-reaction of the M1/70 antibody with CD11b expressed on human monocytes, polymorphonuclear leukocytes, and NK cells has been reported.
570292 Rev. 1
フォーマットの詳細
RB705
The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
RB705
Blue 488 nm
498 nm
707 nm
570292 Rev.1
引用＆参考文献
Development References (12)
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Ault KA, Springer TA. Cross-reaction of a rat-anti-mouse phagocyte-specific monoclonal antibody (anti-Mac-1) with human monocytes and natural killer cells. J Immunol. 1981; 126(1):359-364. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting, Radioimmunoassay). View Reference
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Beller DI, Springer TA, Schreiber RD. Anti-Mac-1 selectively inhibits the mouse and human type three complement receptor. J Exp Med. 1982; 156(4):1000-1009. (Clone-specific: Blocking, Inhibition). View Reference
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Driver DJ, McHeyzer-Williams LJ, Cool M, Stetson DB, McHeyzer-Williams MG. Development and maintenance of a B220- memory B cell compartment. J Immunol. 2001; 167(3):1393-1405. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
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Kaji K, Takeshita S, Miyake K, Takai T, Kudo A. Functional association of CD9 with the Fc gamma receptors in macrophages. J Immunol. 2001; 166(5):3256-3265. (Clone-specific: Fluorescence microscopy, Immunofluorescence). View Reference
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Kishimoto TK, Jutila MA, Berg EL, Butcher EC. Neutrophil Mac-1 and MEL-14 adhesion proteins inversely regulated by chemotactic factors. Science. 1989; 245(4923):1238-1241. (Biology). View Reference
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Lagasse E, Weissman IL. Flow cytometric identification of murine neutrophils and monocytes. J Immunol Methods. 1996; 197(1-2):139-150. (Clone-specific: Flow cytometry). View Reference
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Lub M, van Kooyk Y, Figdor CG. Competition between lymphocyte function-associated antigen 1 (CD11a/CD18) and Mac-1 (CD11b/CD18) for binding to intercellular adhesion molecule-1 (CD54). J Leukoc Biol. 1996; 59(5):648-655. (Clone-specific: Immunoprecipitation). View Reference
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Sanchez-Madrid F, Simon P, Thompson S, Springer TA. Mapping of antigenic and functional epitopes on the alpha- and beta-subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and Mac-1. J Exp Med. 1983; 158(2):586-602. (Clone-specific: Immunoaffinity chromatography, Immunoprecipitation, Inhibition). View Reference
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Springer T, Galfre G, Secher D, Milstein C. Monoclonal xenogeneic antibodies to mouse leukocyte antigens: identification of macrophage-specific and other differentiation antigens. Curr Top Microbiol Immunol. 1978; 81:45-50. (Immunogen: Immunoprecipitation, Radioimmunoassay). View Reference
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Springer T, Galfre G, Secher DS, Milstein C. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody. Eur J Immunol. 1979; 9(4):301-306. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
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Springer T, Galfre G, Secher DS, Milstein C. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur J Immunol. 1978; 8(8):539-551. (Immunogen: Immunoprecipitation). View Reference
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Springer TA, Davignon D, Ho MK, Kurzinger K, Martz E, Sanchez-Madrid F. LFA-1 and Lyt-2,3, molecules associated with T lymphocyte-mediated killing; and Mac-1, an LFA-1 homologue associated with complement receptor function. Immunol Rev. 1982; 68:171-195. (Immunogen: Immunoprecipitation, Radioimmunoassay). View Reference
570292 Rev. 1
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.
