mult1 monoclonal antibody  (Thermo Fisher)

Journal: Journal of Inflammation Research

Article Title: MULT1-Encoding DNA Alleviates Schistosomiasis-Associated Hepatic Fibrosis via Modulating Cellular Immune Response

doi: 10.2147/JIR.S354224

Figure Lengend Snippet: MULT1 encoding DNA injection alleviates egg granuloma and hepatic fibrosis in mice infected with Schistosoma japonicum . ( A ) Experimental design. Briefly, 6-week-old BALB/c female mice (n=6 per group) were artificially infected with S. japonicum by cutaneous contact with 24 cercariae on a wet cover slip. Administration of 40 μg rMULT1 DNA or vehicle DNA was carried out via hydrodynamic tail vein injection; the process was initialized at 4 weeks post infection and repeated 3 times in 1 month before the mice were anaesthetized and sacrificed at the end point. ( B ) RT-qPCR data and ( C ) sandwich fluorescence immunoassay showing elevated MULT1 expression in p-rMULT1-injected mice compared with control group mice (GFP-ctl) administered vehicle plasmids. ( D ) Representative H&E staining images (left panel, magnification x200) and quantification of mean (±SEM) egg-induced granuloma size in the liver (white circles). ( E ) Representative Masson’s trichrome staining images (left panel, magnification x200) and quantification of mean (±SEM) collagen deposition (right panel). ( F and G ) RT-qPCR showing decrease of ( F ) liver collagen I and ( G ) α-SMA expression. Western blotting assay demonstrating reduced protein concentration of ( H and I ) collagen I, ( H and J ) α-SMA and ( H and K ) TGF-β in the livers of mice administered rMULT1 DNA. Data are representative of 4–6 animals per subgroup and 3 independent experiments. Comparisons were between rMULT1 and GFP-ctl, *P<0.05 and **P<0.01.

Article Snippet: Briefly, nonionic latex beads (Invitrogen; Thermo Fisher Scientific, Inc.) were coated with 25 μg/mL MULT1 monoclonal antibody (clone 5D10; eBioscience; Thermo Fisher Scientific, Inc.) and blocked with 2% bovine serum album and then incubated with liver tissue lysates and then captured MULT1 was detected by staining with fluorescence labeled detecting antibody specific for MULT1 (clone 237104, R&D Systems, Inc.) and analyzing on FACSVerse or LSRII flow cytometer.

Techniques: Injection, Infection, Quantitative RT-PCR, Fluorescence, Expressing, Staining, Western Blot, Protein Concentration

Journal:

Article Title: Costimulation of dendritic epidermal ?? T cells by a new NKG2D ligand expressed specifically in the skin

doi: 10.4049/jimmunol.0802439

Figure Lengend Snippet: Expression of H60c mRNA in tissues and in tumor cell lines. A and B, Total RNA (30 μg) from the indicated tissues (A) or cell lines (B) was electrophoresed on agarose gels and immobilized on a charged nylon membrane. The membrane was hybridized with a 32P-labeled H60c probe (top panel). After the final exposure, the membranes were stripped and rehybridized with a GAPDH probe to control for loading of all lanes (middle panel). Ethidium bromide staining of ribosomal RNA in the gel lanes prior to transfer is shown at the bottom of panel A. As determined in other studies ((25), unpublished data), most of the cell lines examined in panel B expressed one or more of the other known NKG2D ligands (Rae1 (R), Mult1 (M) and/or H60a (A)), including Fibroblast (R, M), Nobo-1 (R, M), Yac-1 (R, M, A), SM-1 (R, M), RMA (no ligands), Tramp-C1 (R), DC2.1 (R, M), and B16-BL6 (no ligands). The results are representative of three experiments.

Article Snippet: The following antibodies specific for NKG2D ligands were purchased from R&D Systems: pan-Rae1 monoclonal antibody (186107), MULT1 monoclonal antibody (237104), and biotinylated H60a polyclonal antibody (Cat# BAF1155).

Techniques: Expressing, Membrane, Labeling, Staining

Journal:

Article Title: Costimulation of dendritic epidermal ?? T cells by a new NKG2D ligand expressed specifically in the skin

doi: 10.4049/jimmunol.0802439

Figure Lengend Snippet: Cultured primary keratinocytes express H60c and activate DETC. A, Dissociated C57BL/6 skin cells were cultured for the indicated period before analysis by flow cytometry. Cells were stained with antibodies that recognize Rae1, MULT1, or H60c (open histogram) or control antibodies (shaded histogram). B, Two color contour plot shows H60c and β4-integrin expression on ex vivo keratinocytes (18-hours post dissociation). C, Increased H60c transcripts in keratinocytes after in vitro culture for 18 hours, determined by quantitative RT-PCR. The data were normalized to GAPDH transcript amounts. D, Target cells used in panel E were stained with antibodies that recognize Rae1, MULT1, or H60c or with NKG2D tetramers (open histograms) or control antibodies or streptavidin (shaded histograms). E, In vitro expanded DETCs from wild-type or Klrk1−/− mice were used as effector cells for cytotoxicity assays against keratinocytes harvested from primary cultures. NKG2D antibody (MI-6) or isotype control antibody was used to block interactions. Keratinocytes were in culture for 30 days. The results are representative of three or more experiments.

Article Snippet: The following antibodies specific for NKG2D ligands were purchased from R&D Systems: pan-Rae1 monoclonal antibody (186107), MULT1 monoclonal antibody (237104), and biotinylated H60a polyclonal antibody (Cat# BAF1155).

Techniques: Cell Culture, Flow Cytometry, Staining, Expressing, Ex Vivo, In Vitro, Quantitative RT-PCR, Blocking Assay