14-0116-80

antibody from Invitrogen Antibodies

Targeting: ITGAX CD11C

Provider product page for 14-0116-80

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [23]
Comments [0]
Validations
Flow cytometry [1]
Other assay [12]

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Product number: 14-0116-80 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD11c Monoclonal Antibody (3.9), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The 3.9 monoclonal antibody reacts with human CD11c, the 150 kDa integrin alphaX chain. CD11c non-covalently associates with beta2 integrin to form the CD11c/CD18 heterodimer. This complex is expressed on monocytes, granulocytes, macrophages, NK, dendritic cells and subset of T and B lymphocytes. CD11c/CD18 binds to CD54, iC3b and fibrinogen and plays a role in leukocyte adhesive interactions. Applications Reported: The 3.9 antibody has been reported for use in flow cytometric analysis, and immunohistochemical staining of frozen tissue sections. Applications Tested: The 3.9 antibody has been tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 3.9
Vial size: 25 µg
Concentration: 0.5 mg/mL
Storage: 4° C

ITGAX protein structure - 14-0116-80 shown in red.

Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Immune profiling in DENV-treated whole blood cells 24 h postincubation. Following DENV (MOI = 1) coculture in 100 mu l of WB ex vivo for 24 h, (a) representative flow cytometric analysis and gating of various cells obtained from five cases, performed by staining for specific cell surface markers (CD4, CD8, CD11c, CD14, CD16, CD19, CD25, CD56, CD62L, and HLA-DR), in the DENV-infected and mock groups showed (b) the changes in the expression of specific immune cell populations as noted. (c) The results are shown as a percentage of the mean +- SD obtained from five cases. * p < 0.05, ** p < 0.01, and *** p < 0.001, compared to the mock group. R: region; WBC: white blood cell; bri: bright.

Supportive validation

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Experimental details: Figure 1. Gating Strategy to Sort Human and Mouse Mononuclear Phagocytes for RNA Isolation (A and B) Fluorescence-activated cell sorting (FACS) gating strategy used to isolate human (A) or mouse (B) mononuclear phagocytes (MPs, labeled in red) from the indicated tissues. Before sorting, cell suspensions were magnetically enriched as indicated and gated to select Live; Single; CD45 + ; Lineage - cells as shown in Figures S1 and S2 . Arrows indicate where further analysis is performed on the specified subpopulation. The data presented are representative of 3-6 replicates per sort. (C) Table illustrating cell surface marker expression by MP subsets in human (top) and mouse (bottom). - or lo signifies no or little expression, + or ++ indicates expression or strong expression, -/+ indicates heterogeneous expression of marker proteins.

Supportive validation

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Experimental details: FIGURE 1 Extraction, identification, and purification of myeloid-derived suppressor cell and cultivation of dendritic cell. (A) Cells were extracted from the bone marrow of BALB/c mice and stained by monoclonal antibodies. Under flow cytometry, the expressive rate of Gr-1 + MDSC, CD11b + MDSC, CD11c + MDSC, CD80 + MDSC, F4/80 + MDSC, and MHC-II + MDSC were 70.4%, 3.5%, 4.8%, 1.2%, 0.3%, 2.1% respectively. (B) The expressive rate of Gr-1 + CD11b + MDSC was 22.6%. (C) After MACS by CD11b magnetic bead, purification of Gr-1 + CD11b + MDSC reached 84.6%. (D) Most non-antigen-loaded dendritic cells grew adherently, with different sizes, star or spindle shape, and stretching tubers, but some of the cells seemed to have adopted a half-adherent state with rough surface. The expressive rates of CD11c, CD86, and MHC-II on DCs were 10.9%, 3.8%, and 27.9%, respectively, by flow cytometry. (E) On the 7th day, DCs were stimulated and activated by tumor antigens. DCs in the half-adherent state increased obviously with radial spikes and bigger shape. The expressive rates of CD11c, CD86, and MHC-II were 74.8%, 50.3%, and 49.8%, respectively, by flow cytometry. IgG FITC, a homotypic control antibody, was used to set the gate strategy. Scale bar = 100 mumol/liter. MDSC, myeloid-derived suppressor cell; MACS, magnetic-activated cell sorting; DC, dendritic cell; CD, cluster of differentiation

Supportive validation

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Experimental details: 5 FIGURE Impact of particle size on aluminum adjuvant internalization. (a) U-937 cells were incubated with AlOOH-lumo (control) or AlOOH-lumo previously subjected to freeze/thaw cycles (freeze/thaw) to increase the size of the particles, as described in experimental procedures. The cells were incubated at 37degC for 4 h and subsequently processed for flow cytometry analysis. Differentiated macrophages were classified using anti-CD11c antibodies. Cell nuclei were labeled with DAPI. Cell populations encircled by a dotted line have not internalized AlOOH-Lumo and cells that have internalized AlOOH-Lumo are encircled by a solid line. (b) U-937 cells were incubated with control or freeze/thaw AlOOH-lumo preparations as described above and subsequently fixed and F-Actin stained with blue phalloidin (pseudocolored gray). Red arrowheads in the right panel point to small, internalized adjuvant particles. Spinning disk confocal images represent a merge of z-stacks. Images are representative of three independent trials. Fifty cells per trial per condition were analyzed. Scale bars, 5 mum. (c) U-937 cells were incubated with control or freeze/thaw AlOOH-lumo preparations and processed as described in (b). The percentage of cells with intracellular AlOOH particles were calculated for three independent trials and the mean represented in the bar graph. * p

Supportive validation

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Experimental details: IsoLG adducts are enriched in monocytes of patients with SLE. ( A ) Representative FACS plots displaying isoLG adduct containing CD11c + PBMCs from a representative control and patient with SLE. Representative histograms displaying the distribution of isoLG adducts in ( B ) CD11c + and ( C ) CD11c + CD86 + cells. Quantitation of IsoLG adduct-containing cells as a percentage of ( D ) CD11c + , ( E ) CD11c + CD86 + , and ( F ) CD14 + cells. For B - F data were analyzed using 1-tailed Student's t test or Mann-Whitney U test ( n = 10-11, *P < 0.05). ( G ) Stable isotope dilution multiple reaction monitoring for mass spectrometry analysis of isoLG-lysine-lactam adduct in DCs. Representative liquid chromatography/mass spectrometry chromatographs from a representative patient. The top row shows multiple reaction monitoring chromatographs for isoLG lysine lactam in samples, while the bottom row shows multiple reaction monitoring chromatograph for [13C615N2] internal standard for the same samples. cps, counts per second; Rt, retention time. ( H ) Quantitation of isoLG-lysine in monocytes from a subset of SLE patients and controls. ( I ) Monocytes from SLE patients and controls were sorted. Superoxide was detected using HPLC to monitor conversion of dihydroethidium to the superoxide oxidation adduct 2-hydroxyethidium (2-HO-ET) and ethidium. ( J ) Quantitation of 2-HO-ET from SLE patients and controls. For H and J , comparisons were made with a 1-tailed Student's t test ( n = 4-6, *P

Supportive validation

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Experimental details: Figure 2 Protein delivery efficiency of dNP2 in primary mouse and human immune cells. ( a , b ) Mouse primary splenocytes were isolated from 6-week-old female C57BL/6 mice and the cells were incubated with 5 muM EGFP, TAT- and dNP2-EGFP for 2 h. Intracellular fluorescence was analysed by flow cytometry and the data are represented as dot plots or mean fluorescence intensity (MFI) of the cells. ( c , d ) Human PBMCs were isolated from healthy donor blood and the cells were incubated with 5 muM EGFP, TAT-, dNP2-EGFP for 2 h. The data were analysed as described above. ( e ) Total splenocytes were incubated with 1 muM EGFP, TAT-, and dNP2-EGFP for 2 h. Cells were gated using markers specific for CD4 T cells (CD4 + ), B cells (CD19 + ), macrophages (CD11c lo CD11b hi F480 + ) and DCs (CD11c hi MHCII hi ). The EGFP signal in each cell population was then analysed by flow cytometric analysis. The relative MFI value was normalization to PBS treated cells. The red line indicates relative MFI of PBS-treated cells. ( f ) Total PBMCs were incubated with 1 muM EGFP, TAT-, and dNP2-EGFP for 2 h. Cells were gated with markers specific for CD4 T cells (CD4 + ), B cells (CD19 + ), macrophages (CD11b + ) and DCs (CD11c + ) and the data were then analysed as described above. ( g ) Time-lapse images of mouse CD4 T cells incubated with 1 muM EGFP, TAT- and dNP2-EGFP were acquired for 2 h (Scale bar, 15 mum) and ( h ) the average fluorescence intensities of 10 cells from each sample were calculate

Supportive validation

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Experimental details: Figure 2. Percentage of purified DC samples in all experiments. DCs were labeled with the Non-DC Depletion Cocktail for negative selection. Next, DCs were labeled with DC Enrichment Cocktail for positive selection. The expression of CD11c and CD86 was tested by flow cytometery. The percentages of purified DC are presented in the top of each panel. DCs, dendritic cells; CD, cluster of differentiation.

Supportive validation

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Experimental details: Figure 2. Detection of surface markers CD11c and CD206 expression was carried out using flow cytometry approach in the synovial fluids biopsied and peripheral blood from participants. (A) Expression of CD11c and CD206 surface markers of M1 and M2 macrophages was evaluated using flow cytometry in the synovial fluids biopsied from 35 patients with knee OA and paired healthy control. (B) Analysis of ratio of M1 to M2 macrophages in healthy controls and knee OA. (C) Likewise, expression of CD11c and CD206 surface markers was assayed using flow cytometry in the peripheral blood sampled from 80 cases of patients with knee OA and paired healthy control. (D) Similarly, ratio of M1 to M2 macrophages was calculated in healthy controls and knee OA. In light of the data being non-normal distribution, Mann-Whiney U test was used to analyze the statistical difference between healthy control and knee OA. *P

Supportive validation

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Experimental details: Figure 1. CASC15 -KO promotes the differentiation of acute myeloid leukemia cells. (A) Apoptosis in CASC15 -KO and empty vector-transduced (control) OCI-AML5 cell lines after 24 h of depletion of granulocyte-macrophage colony-stimulating factor (annexin-FITC/Sytox blue flow cytometry). (B) Expression of SOX4 during in vitro differentiation of CASC15 -KO and control OCI-AML5 cell lines. All cells were treated with 0.1 mM all- trans retinoic acid (ATRA) and 1 nM 12-O-tetradecanoylphorbol-13-acetate (TPA) over 72 h in three independent experiments. Total RNA was extracted before, after 24 h and after 72 h of treatment, DNase-digested and transcribed to cDNA. A quantitative real-time polymerase chain reaction (qRT-PCR) was performed using SYBR green chemistry with subsequent melting curve analysis in technical triplicates. The 2-ddCt was calculated relative to the pre-determined housekeeping gene encoding succinate dehydrogenase complex subunit C ( SDHC ). (C) Baseline expression of the monocyte/macrophage markers CD11b (integrin subunit alpha M, ITGAM), CD11c (integrin subunit alpha X, ITGAX), and CD14, the granulocyte marker CD15 (fucosyltransferase 4, FUT4), and the general myeloid marker CD13 (aminopeptidase N, APN) in CASC15 -KO and control cells. The percentages of positive cells, quantified by flow cytometry after 72 h, are shown. (D-F) Growth rate and CD11c myeloid cell surface marker expression of CASC15 and control cell lines during drug-induced in vitro differentiation