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R718 Rat Anti-CD11b
R718 Rat Anti-CD11b
Multiparameter flow cytometric analysis of CD11b expression on mouse bone-marrow cells. 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™ R718 Rat IgG2b, κ Isotype Control (Cat. No. 566943; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD11b antibody (Cat. No. 567469; Right Plot) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of CD11b (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 cell populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
R718 Rat Anti-CD11b
Multiparameter flow cytometric analysis of CD11b expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ R718 Rat IgG2b, κ Isotype Control (Cat. No. 566943; Left Plot) or BD Horizon™ R718 Rat Anti-CD11b antibody (Cat. No. 567469; Right Plot) at 0.5 µg/test. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD11b (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 intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD11b expression on mouse bone-marrow cells. 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™ R718 Rat IgG2b, κ Isotype Control (Cat. No. 566943; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD11b antibody (Cat. No. 567469; Right Plot) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of CD11b (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 cell populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD11b expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ R718 Rat IgG2b, κ Isotype Control (Cat. No. 566943; Left Plot) or BD Horizon™ R718 Rat Anti-CD11b antibody (Cat. No. 567469; Right Plot) at 0.5 µg/test. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD11b (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 intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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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
16409
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 CompBead 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 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.
  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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  7. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
567469 Rev. 1
Antibody Details
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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.

  

The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628–640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

567469 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
567469 Rev.1
Citations & References
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View product citations for antibody "567469" on CiteAb

Development References (11)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
View All (11) View Less
567469 Rev. 1

 

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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.