CD11b Antibody | CA16.3E10

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Immunophenotyping gating strategy and morphological analysis for MDSC identification in peripheral blood of dogs. PBMCs from healthy dogs and dogs with cancer were stained for the myeloid marker CD11b, monocytic marker CD14 and MHC II. (A) Representative flow cytometric analysis of forward and side scatter and gated CD11b+CD14-MHCII- cells from dogs with advanced or metastatic tumors compared to dogs with early stage non-metastatic tumors and healthy control dogs. Plots are representative of dog with advanced metastatic hemangiosarcoma (top), early stage bladder transitional cell carcinoma (middle) and a healthy dog. (B) FACS sorted CD11b+CD14-MHCII- cells were stained with diff-quick for cell morphology evaluation. A representative example of polymorphonuclear granulocyte morphology of CD11b+CD14-MHCII- cells is shown at 63× magnification.

From: Goulart MR, Pluhar GE, Ohlfest JR (2012) Identification of Myeloid Derived Suppressor Cells in Dogs with Naturally Occurring Cancer. PLoS ONE 7(3): e33274.

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CD11b+CD14-MHCII- cells suppress T cell proliferation. Facs sorted CD11b+CD14-MHCII- cells isolated from a dog with osteosarcoma or healthy PBMCs were co-incubated with mitogen-stimulated CD4+ and CD8+ T cells isolated from a healthy dog for 72 hs. No stimulated cells were used as negative control. Proliferative responses were measured by 3H-thymidine incorporation from experiments performed in triplicate. CPM, counts per minute. Mean ± SEM are shown.

From: Goulart MR, Pluhar GE, Ohlfest JR (2012) Identification of Myeloid Derived Suppressor Cells in Dogs with Naturally Occurring Cancer. PLoS ONE 7(3): e33274.

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Mouse anti-CD11b and Gr-1 antibodies cross-react with canine samples. Fresh PBMCs from healthy dog and cancer patients were isolated by Ficoll, stained with anti-mouse CD11b and anti-mouse Gr-1 antibodies.

From: Goulart MR, Pluhar GE, Ohlfest JR (2012) Identification of Myeloid Derived Suppressor Cells in Dogs with Naturally Occurring Cancer. PLoS ONE 7(3): e33274.

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CD11b+CD14+MHCII- cells demonstrate ability to suppressive T cell proliferation. (A) CD11b+CD14+MHCII- cells were sorted from peripheral blood sample of an osteosarcoma dog (B) and co-cultured with healthy dog PBMCs in the presence of mitogen for 72 hs. Non-stimulated PBMCs were used as negative control and PBMCs co-cultured with healthy PMNs were used to control for the effect of adding cells to the assay. Proliferative responses were measured by 3H-thymidine incorporation. CPM, counts per minute. The experiment was performed in triplicate. Mean ± SEM are shown.

From: Goulart MR, Pluhar GE, Ohlfest JR (2012) Identification of Myeloid Derived Suppressor Cells in Dogs with Naturally Occurring Cancer. PLoS ONE 7(3): e33274.

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Flow cytometer analysis of CD11b integrin expression and the binding of Leishmania promastigotes to canine monocytes. (A) Gate R1 was built separating monocytes from animals naturally infected (ANI) with L. chagasi by cells size (x axis) and granularity (y axis). (B) Gate with mononuclear cells (control) (C) CD11b positive cells; (D) CD11b expression during the L. chagasi binding to mononuclear cells with the presence of C5D serum. (E) CD11b expression during the L. chagasi binding to mononuclear cells with the absence of C5D serum.

From: Sampaio et al. BMC Veterinary Research 2007 3:11

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Staining characteristics and evaluation of two commercially available canine granulocytic antibodies of canine peripheral blood. (a) Cells were analyzed according to forward and side scatter. Cells were either gated to include all live and dead (no gate), all live cells including lymphocytes (R1, lymphocytes are circled) and all myeloid cells (P1). (b) Cells were labeled using canine anti-CD11b and either (a) CADO48A or (b) DH59B with the same concentration of secondary antibody FITC. As can be seen in (a), CADO48A allows visualization of an additional cell population as compared to DH59B (b). These results were repeatable with multiple blood samples from different dogs. The cells in (b) were gated on P1 as indicated in (a).

From: Sherger et al. BMC Veterinary Research 2012 8:209

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Optimization of secondary antibody staining concentration for detection of CADO48A. Cells were stained with the primary antibody CADO48A alone followed by (a) 1:10 (b) 1:50 and (c) 1:100 dilutions of a secondary FITC antibody. Optimal detection was seen between 1:50 and 1:100 dilution with CADO48A alone. Secondary FITC concentrations of 1:50 (d) and 1:100 (e) to detect CADO48A on pre-labeled CD11b cells were then evaluated and demonstrated that a 1:50 concentration of FITC was optimal for optimal detection of distinct CADO48A+ cells. Based on these findings, a 1:50 FITC concentration was used in all clinical samples. All cells in these diagrams were gated on P1.

From: Sherger et al. BMC Veterinary Research 2012 8:209

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Effects of EDTA versus media storage of patient blood samples on flow cytometric results. Peripheral blood cells were (a) stained immediate following collection with CD11b and CADO48A or maintained in EDTA collection tubes for (b) 24 or (c) 48 hours or in media for (d) 24 or (e) 48 hours prior to staining with CD11b and CADO48A. Both EDTA and media samples were kept refrigerated. These findings show a decrease over time of population distinction in EDTA with minimal changes when cells are kept in media up to 48 hours. All cells were gated on P1.

From: Sherger et al. BMC Veterinary Research 2012 8:209

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Effects of fixation on CD11b and CADO48A expression levels. Cells were stained for CD11b and CADO48A and evaluated pre-fixation (a) or subsequently fixed with 4% paraformaldehyde at (b) 4 hours, (c) 24 hours), (d) 48 hours or (e) 72 hours post-fixation before analysis on the flow cytometer. Fixation of cells appears to have limited effects on expression levels of CD11b and CADO48A expression up to 24 hours and then only mild increases of 4-6% of CD11b+CADO48A+ cells are seen at 24, 48 and 72 hours post-fixation. All cells seen in this diagram were gated on P1.

From: Sherger et al. BMC Veterinary Research 2012 8:209

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Gated myeloid subpopulations evaluated for patient population. All peripheral blood myeloid cells were evaluated using the P1 gate (non-lymphocytes) and subsequently gated based on relatively expression levels of CD11b and CADO48A where CADO48Ahi/CD11bhi (gate P2), CADO48Alow/CD11bhi (gate P3) and CADO48Alow/CD11blow (gate P4) likely represent granulocytes/neutrophils, monocytes and MDSCs, respectively.

From: Sherger et al. BMC Veterinary Research 2012 8:209

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Increased percentage of CD11blow/CADO48Alow in tumor-bearing canine patients. The expression levels of CD11blow and CADO48Alow was evaluated from the peripheral blood of representative canine patients and show an increase of CD11blow/CADO48low in tumor-bearing canine patients.

From: Sherger et al. BMC Veterinary Research 2012 8:209

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Representative gating strategies demonstrating fluorescence profiles of cells in jejunum and colon lamina propria cells in dogs naturally infected with Leishmania; (A) macrophages were identified on the basis of their specific forward (FSC) and side (SSC) light-scattering properties (B) isotype control cutoff; (C) frequency of TLR2+; (D) frequency of TLR9+; (E) frequency of CD14+; (F) frequency of CD11c+; (G) frequency of CD11b+; (H) frequency of CD11b+/CD14+; (I) frequency of TLR9+/CD11b+; (J) frequency of TLR9+/CD14+.

From: Figueiredo MM, Amorim IF, Pinto AJ, Barbosa VS, Pinheiro Lde J, Deoti B, Faria AM, Tafuri WL. Expression of Toll-like receptors 2 and 9 in cells of dog jejunum and colon naturally infected with Leishmania infantum. BMC Immunol. 2013 May 14;14:22.

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Figure A. FITC conjugated Mouse anti Canine CD45 (MCA1042F) and purified Mouse IgG1 isotype control (MCA928) detected with Goat anti Mouse IgG1 DyLight 649 (STAR74D649). Figure B. FITC conjugated Mouse anti Canine CD45 (MCA1042F) and purified Mouse anti Canine CD11b (MCA1777S) detected with Goat anti Mouse IgG1 DyLight 649 (STAR74D649). All experiments performed on red cell lysed canine blood gated on mononuclear cells.

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  • Mouse anti Dog CD11b
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    CA16.3E10
  • Isotype
    IgG1
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA1777SC *, F, IPdatasheet pdfdatasheet pdf2 ml
    MCA1777S
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Mouse anti Dog CD11b antibody, clone CA16.3E10 is a monoclonal antibody recognising the canine CD11b cell surface antigen, a member of the alpha integrin family. CD11b forms one of the possible alpha chains of the canine leukocyte adhesion complexes (LeuCAMs), these contain a common 95 kDa β chain (CD18) non-covalently bound to either a 150 kDa (CD11c), 165 kDa (CD11b) or 180 kDa (CD11a) α chain (Moore et al. 1990. The CD11/CD18 complex is also known as the CR3 receptor.

      Canine CD11b is expressed by granulocytes, monocytes, NK cells and some macrophages. Mouse anti Dog CD11b (CA16.3E10) has been used tio evaluate the effect of anaesthetic administration of CD11b expression on canine neutrophils (Maeda et al. 2010) demonstrating attenuation of CD11b expression at high concentrations administered lidocaine hydrochloride and reduced adhesion of neutrophils to endothelium.
    • Intended Use
    • Target Species
      Dog
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      Goatyes
      Catyes
      Mustelidyes
      Pigyes
      Bovineyes
      Minkyes
      Beluga whaleyes
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      Tissue Culture Supernatant - liquid
    • Reconstitution
    • Preparation
    • Preservative Stabilisers
      0.1%Sodium Azide
    • Immunogen
      Affinity purified beta-2 integrins from splenic lysate
    • Purity
    • Approx. Protein Concentrations
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Storage
      Store at +4oC or at -20oC if preferred.

      This product should be stored undiluted.

      Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      18 months from date of despatch.
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      Flow CytometryNeat
      Immunohistology - Paraffin
      Immunoprecipitation
      Immunohistology - Frozen(1)
      (1)
      The epitope recognised by this antibody is reported to be sensitive to formaldehyde fixation and tissue processing. Bio-Rad recommends the use of acetone fixation for frozen sections.

    • Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested recommended dilutions are given as guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • Flow Cytometry
      Use 10ul of the suggested working dilution to label 106 cells or 100ul whole blood
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional CD11b Antibody Formats

    Formats Clone Applications Sizes available
    CD11b Antibody : S/N CA16.3E10 C *, F, IP 2 ml
    • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

    Recommended Secondary Antibody

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      STAR117D800GA
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      STAR8D800GA
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      STAR117F
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      STAR70
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      STAR9B
      Human anti Mouse IgG1:HRPHCA036P0.1 mgE
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      Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed)STAR117P0.5 mgE, WB
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      Goat anti Mouse IgG:HRP (Rat Adsorbed)STAR770.5 mgC, E, P
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      Goat anti Mouse IgG (Fc):HRPSTAR120P1 mgE, WB
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      Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)STAR13B1 mgC, E, P, RE, WB
      STAR13B
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      STAR76

      Recommended Negative Isotype Control

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse IgG1 Negative ControlMCA928100 TestsF
        MCA928

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              • Application NameReference Images
                Immunofluorescence
                Immunohistology - Frozen

              References

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