Published customer image: 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
Published customer image: 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.
Published customer image: 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
Published customer image: 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
Published customer image: 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.
Published customer image: 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 63x 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.
Published customer image: 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
Published customer image: 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.
Published customer image: 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
Published customer image: 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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