Human TGF-beta  RII Fluorescein-conjugated Antibody

Detection of TGF-beta RII in NS0 Mouse Cell Line Transfected with TGF-beta RII by Flow Cytometry.

NS0 mouse myeloma cell line transfected with TGF-beta RII was stained with Mouse Anti-Human TGF-beta RII Fluorescein-conjugated Monoclonal Antibody (Catalog # FAB241F, filled histogram) or isotype control antibody (Catalog # IC002F, open histogram). View our protocol for Staining Membrane-associated Proteins.

Detection of Human TGF-beta RII by Immunocytochemistry/Immunofluorescence

Investigating the status of the canonical TGF-beta pathway and levels of Hsp90 in the genetically paired SW480 primary and SW620 secondary tumour-derived colon cancer cell lines. a The secretion of TGF-beta 1 by SW480 and SW620 cells was determined by means of an ELISA to quantify the levels of extracellular TGF-beta 1 in spent culture medium for a given cell number after 12 h incubation. Data represent the average of three independent experiments performed in triplicate and error bars indicate the standard error in the mean. b The levels of TGF-beta RII receptor on the cell surface was determined by flow cytometry using a fluorescein isothiocyanate-conjugated antibody. Data was analysed using FlowJo software and gating carried out according to the IgG1 isotype control. Histograms show a shift in fluorescence for each sample (black line, unshaded) compared to the isotype control (grey shading) and the percentage of positive events is indicated for each sample. Data are representative of three independent experiments showing consistent results. c Intracellular levels of TGF-beta 1 in SW480 and SW620 cells were determined by (i) western blot analysis and compared to a histone loading control. The images are cropped from the full length western blot provided as Additional file 1. (ii) The normalized levels of pre-pro TGF beta1 from the western blot in (i) were determined by densitometry using ImageJ and calculated according the histone loading control. AU: arbitrary units. The positive control represents commercially obtained recombinant human TGF beta1 in its active form. d Activation of the TGF-beta canonical pathway was determined by the phosphorylation and nuclear localization of Smad2/3. SW480 and SW620 cells were stained for pSMAD2/3 and nuclei were stained with Hoescht-33342. Immunofluorescence was detected using the Zeiss LSM 510 Meta laser scanning confocal microscope and the images analysed using Axiovision LE 4.7.1 software (Zeiss). (i) The first column shows the level of pSmad2/3 staini