Flow cytometric staining of mouse peripheral blood granulocytes with RPE conjugated Rat anti Mouse Beta-glucan Receptor (MCA2289PE)

Flow cytometric staining of mouse peripheral blood granulocytes with Rat anti Mouse Beta-glucan Receptor: Alexa Fluor®647 (MCA2289A647)

Flow cytometric staining of mouse peripheral blood granulocytes with FITC conjugated Rat anti Mouse Beta-glucan Receptor (MCA2289FA)

Flow cytometric staining of mouse peripheral blood granulocytes with biotin conjugated Rat anti Mouse Beta-glucan Receptor (MCA2289B)

Flow cytometric staining of mouse peripheral blood monocytes with low endotoxin Rat anti Mouse Beta-glucan Receptor (MCA2289EL)

Flow cytometric staining of mouse peritoneal macrophages with FITC conjugated Rat anti Mouse Beta-glucan Receptor (MCA2289FB)

Immunoperoxidase staining of a mouse lymph node cryosection with Rat anti Mouse Dectin-1 antibody, clone 2A11 (MCA2289) followed by horseradish peroxidase conjugated Goat anti Rat IgG antibody (STAR72). Low power

Immunoperoxidase staining of a mouse lymph node cryosection with Rat anti Mouse Dectin-1 antibody, clone 2A11 (MCA2289) followed by horseradish peroxidase conjugated Goat anti Rat IgG antibody (STAR72). Medium power

Immunoperoxidase staining of a mouse skin cryosection with Rat anti Mouse Dectin-1 antibody, clone 2A11 (MCA2289) followed by horseradish peroxidase conjugated Goat anti Rat IgG antibody (STAR72). Low power

Immunoperoxidase staining of a mouse lymph node cryosection with Rat anti Mouse Dectin-1 antibody, clone 2A11 (MCA2289) followed by horseradish peroxidase conjugated Goat anti Rat IgG antibody (STAR72). High power

Published customer image:
Rat ani Mouse Dectin 1 antibody, clone 2A11 (MCA2289) used for the evaluation of Dectin-1 expression on neutrophils by flow cytometry.Image caption:
Ex vivo recognition of yeast particles and live fungi by inflammatory cells. A) Representative flow-cytometric analysis, gated on Ly-6G+ neutrophils, after coincubation of BIOgel-elicited inflammatory cells from wild type or dectin-1-deficient (Clec7a-/-) 129S6/SvEv interaction with serum-opsonized or non-opsonized zymosan. Positive staining for the A405-labelled zymosan identifies the neutrophils that are associated zymosan and only these cells exhibit conversion of APF, the ROS reporter. B) Representsative flow-cytometric analysis of CD11b and dectin-1 expression by inflammatory neutrophils and monocyte/Mø. Data represents specific receptor staining (shaded histograms) and isotype control staining (bold lines). Data representative of 2 independent experiments and consistent with previous experiments with thioglycollate. C) Inflammatory cells were loaded with APF and then incubated with serum-opsonized or non-opsonized A405-labelled zymosan or Pacific Orange-labelled C. albicans for 15 minutes. After this time the association of the inflammatory cells with zymosan was measured by flow-cytometry (upper panels) and in those cells that were interacting with zymosan the evidence for fluorescent conversion of APF was also quantified (lower panels). Data is derived from three independent experiments and the data derived from the use of dectin-1-deficient cells is shown relative to wild type cells (100%) as mean±95% confidence interval (raw representative data from one of the 3 independent experiments are shown in the Figure S1). The impact of complement osponization (‘C’) and the use of different fungal particle used (‘F’) were assessed by Two-way ANOVA (‘I’ = Interaction statistic). Samples in which the 95% confidence intervals do not overlap with the mean wildtype are specific indicated with a # symbol. Differences in impairment of response observed with dectin-1-deficient cells were further analysed by Bonferroni post-tests. P values derived from individual Bonferroni post-tests are indicated with bracketed pairs of samples.

From: McDonald JU, Rosas M, Brown GD, Jones SA, Taylor PR (2012) Differential Dependencies of Monocytes and Neutrophils on Dectin-1, Dectin-2 and Complement for the Recognition of Fungal Particles in Inflammation. PLoS ONE 7(9): e45781

Published customer image:
Rat anti Mouse dectin-1 antibody, clone 2A11 (MCA2289) used for the evaluation of dectin-1 expression on wild type and knockout mouse macrophages by flow cytometry.
Image caption:
Dectin-1 response and expression is Ae2-dependent.
(A and B) Tnfα mRNA expression relative to Hprt in Ae2wt/wt and Ae2Δ/Δ macrophages stimulated for 6h with Zymosan (5 ppc) (A) or PAM3CK4 (B) (n = 8–10, ** p<0.01, Mann-Whitney test). (C) Ae2 mRNA expression relative to Hprt by WT macrophages stimulated for 6h with Zymosan (Zym, 5 ppc) or PAM3CK4 (100 ng/ml) (n = 8–12, * p<0.05, *** p<0.001, Mann-Whitney test). (D) Representative flow cytometry histogram with quantification by mean fluorescence intensity (MFI) of Dectin-1 expression on cell surface of Ae2wt/wt and Ae2Δ/Δ macrophages (n = 8, *** p<0.001, Mann-Whitney test). (E) Representative flow cytometry histogram and MFI of CD64 expression on cell surface of Ae2wt/wt and Ae2Δ/Δ macrophages (n = 5).

From: Urso K, Charles JF, Shull GE, Aliprantis AO, Balestrieri B (2016)
Anion Exchanger 2 Regulates Dectin-1-Dependent Phagocytosis and Killing of Candida albicans.
PLoS ONE 11(7): e0158893.

Published customer image:
Rat anti Mouse dectin-1 antibody, clone 2A11 (MCA2289) used for the evaluation of dectin-1 expression on wild type and treated mouse macrophages by flow cytometry.
Image caption:
Pharmacological alkalinization of pHi inhibits Dectin-1 expression.
(A) Dectin-1 expression relative to Hprt by wild-type macrophages treated with bafilomycin A (Baf A, 25 nM) or DMSO control for 6 and 16 h. (B) Representative flow cytometry histogram and (C) quantification by MFI showing decreased Dectin-1 in wild-type macrophages treated with Baf A for 6 and 16 h compared to DMSO control (n = 6, ** p<0.01, Mann-Whitney test).

From: Urso K, Charles JF, Shull GE, Aliprantis AO, Balestrieri B (2016)
Anion Exchanger 2 Regulates Dectin-1-Dependent Phagocytosis and Killing of Candida albicans.
PLoS ONE 11(7): e0158893.

Published customer image:
Rat anti Mouse Dectin-1 antibody, clone 2A11 (MCA2289) used for the evaluation of surface dectin-1 expression by murine peritoneal and bone marrow derived macrophages by flow cytometry.
Image caption:
Murine primary macrophages including peritoneal macrophages and BMDM express similar levels of the BG receptor dectin-1 and exhibit the same ability to respond to Sc BG extracts.
(A-B) Freshly isolated 4-days thioglycollate-elicited peritoneal macrophages (TEPM), resident peritoneal macrophages (RPM) and BMDM from WT and Clec7a^-/- C57Bl/6, and DBA/2 mice, and the macrophages cell line RAW-Blue™, used as control, were assessed for surface expression of dectin-1 by flow cytometry (MACSQuant®, Miltenyi Biotech, Germany). Doublets of cells were excluded at the beginning of the acquisition and a 7-AAD staining was used to discriminate death cells. The surface expression of dectin-1 was determined for each type of primary macrophages on CD115⁺ cells. (A) Data are expressed as the frequency of dectin-1⁺ among CD115⁺ cells. (B) The mean of fluorescence intensity (MFI) of dectin-1 was established on CD115⁺ cells. Results are presented as the mean ± SD MFI of dectin-1 corrected by either the MFI of isotype control for RAW macrophages, or the MFI of Clec7a^-/- C57Bl/6 for WT C57Bl/6 or DBA/2 primary macrophages. (C) Remaining TEPM and BMDM from WT and Clec7a^-/- C57Bl/6 mice isolated from these experiments were subcultured and stimulated with 100 μg/mL of crude or enriched Sc BG CW extracts (BG15, BG65 or BG75) or zymosan for 8 h (37°C, 5% CO₂). Supernatants were collected immediately after incubation and were assessed for TNFα production by ELISA. Results are shown as the mean ± SD. All data are representative of three mice per group from three independent experiments performed in triplicate. *p <0.05 (two-tailed unpaired Student's t-test); mean values not sharing the same letter are significantly different.

From: Walachowski S, Tabouret G, Foucras G (2016) Triggering Dectin-1-Pathway Alone Is Not Sufficient to Induce Cytokine Production by Murine Macrophages. PLoS ONE 11(2): e0148464.