- 67-0116-42 - Invitrogen Antibodies ITGAX antibody

Submitted references Aluminum hydroxide adjuvant diverts the uptake and trafficking of genetically detoxified pertussis toxin to lysosomes in macrophages.

Jaldin-Fincati J, Moussaoui S, Gimenez MC, Ho CY, Lancaster CE, Botelho R, Ausar F, Brookes R, Terebiznik M
Molecular microbiology 2022 May;117(5):1173-1195

Isolevuglandins disrupt PU.1-mediated C1q expression and promote autoimmunity and hypertension in systemic lupus erythematosus.

Patrick DM, de la Visitación N, Krishnan J, Chen W, Ormseth MJ, Stein CM, Davies SS, Amarnath V, Crofford LJ, Williams JM, Zhao S, Smart CD, Dikalov S, Dikalova A, Xiao L, Van Beusecum JP, Ao M, Fogo AB, Kirabo A, Harrison DG
JCI insight 2022 Jul 8;7(13)

Upregulation of CD3ζ and L-selectin in antigen-specific cytotoxic T lymphocytes by eliminating myeloid-derived suppressor cells with doxorubicin to improve killing efficacy of neuroblastoma cells in vitro.

Xu W, Li S, Li M, Zhou H, Yang X
Journal of clinical laboratory analysis 2022 Jan;36(1):e24158

Increased TNF-α Initiates Cytoplasmic Vacuolization in Whole Blood Coculture with Dengue Virus.

Satria RD, Huang TW, Jhan MK, Shen TJ, Tseng PC, Wang YT, Yang ZY, Hsing CH, Lin CF
Journal of immunology research 2021;2021:6654617

The long non-coding RNA Cancer Susceptibility 15 (CASC15) is induced by isocitrate dehydrogenase (IDH) mutations and maintains an immature phenotype in adult acute myeloid leukemia.

Grasedieck S, Ruess C, Krowiorz K, Lux S, Pochert N, Schwarzer A, Klusmann JH, Jongen-Lavrencic M, Herold T, Bullinger L, Pollack JR, Rouhi A, Kuchenbauer F
Haematologica 2020 Sep 1;105(9):e448-453

Human and Mouse Transcriptome Profiling Identifies Cross-Species Homology in Pulmonary and Lymph Node Mononuclear Phagocytes.

Leach SM, Gibbings SL, Tewari AD, Atif SM, Vestal B, Danhorn T, Janssen WJ, Wager TD, Jakubzick CV
Cell reports 2020 Nov 3;33(5):108337

Complex-centric proteome profiling by SEC-SWATH-MS for the parallel detection of hundreds of protein complexes.

Bludau I, Heusel M, Frank M, Rosenberger G, Hafen R, Banaei-Esfahani A, van Drogen A, Collins BC, Gstaiger M, Aebersold R
Nature protocols 2020 Aug;15(8):2341-2386

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Normal human peripheral blood cells were stained with Mouse IgG1 kappa Isotype Control, Super Bright 702 (Product # 67-4714-82) (blue histogram) or CD11c Monoclonal Antibody, Super Bright 702 (purple histogram). Cells in the monocyte gate were used for analysis.

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

Submitted by: Invitrogen Antibodies (provider)
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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

Submitted by: Invitrogen Antibodies (provider)
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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

67-0116-42

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