Journal: Acta Neuropathologica Communications

Article Title: Determinants of astrocytic pathology in stem cell models of primary tauopathies

doi: 10.1186/s40478-023-01655-1

Figure Lengend Snippet: Antibody Product Information

Article Snippet: This was followed by a 30-minute incubation at room temperature with goat anti-mouse IgG H&L Alexa Fluor 555 (ab150114, RRID: AB_2687594; Abcam; Cambridge, UK) and goat anti-rabbit IgG H&L Alexa Fluor 488 (ab150077, RRID: AB_2630356; Abcam; Cambridge, UK) antibodies, both at 1:1000 in TBS + 0.1% BSA.

Techniques: Plasmid Preparation

Journal: Oncotarget

Article Title: Evaluation of a novel human IgG1 anti-claudin3 antibody that specifically recognizes its aberrantly localized antigen in ovarian cancer cells and that is suitable for selective drug delivery

doi:

Figure Lengend Snippet: A. Expression of claudin3 mRNA in human cancer cell lines USPC-4, OSPC-2, UCI-107 and normal human embryonal kidney HEK293 cells was measured by RT-qPCR analysis. Data are shown as the mean ± SE of two independent experiments and are expressed as relative expression ratios (ΔΔCt – Fold increase) using HEK293 as a reference. B. 20 ug of cell lysates were separated by SDS-PAGE and probed with anti-claudin3 polyclonal antibody. Equal loading was confirmed by incubating the membrane with αTubulin. C. Total claudin3 expression level was evaluated by FACS analysis on fixed and permeabilized cells. Single cell suspension were incubated with commercial antibody against claudin3 followed by incubation with anti-rabbit-FITC conjugated antibody. Histograms of cells stained with an isotype control antibody (dotted line) or with specific anti-claudin3 (solid line) are shown for each cell line tested. The percentage of claudin3 positive cells is reported in each plot. D. IgGH6 binding to the extracellular domain of claudin3 was evaluated by FACS analysis on unfixed cells. Single cell suspensions were incubated with 2.5 μg/mL of purified IgGH6 for 1 h followed by a 30 min incubation with mouse anti-human IgG-FITC secondary antibody, and analyzed by FACS gating on annexin V negative cells. Histograms of cells stained with specific IgGH6 (solid line) or only secondary antibody (dotted line) are shown for each of the cell lines tested. The percentage of claudin3 positive cells is reported in each plot.

Article Snippet: After detachment with 0.5 mM EDTA, cells were incubated for 1 h with 2.5 μg/mL of purified IgGH6 followed by a 30 min incubation with mouse anti-human IgG-FITC secondary antibody (Southern Biotech).

Techniques: Expressing, Quantitative RT-PCR, SDS Page, Membrane, Suspension, Incubation, Staining, Control, Binding Assay, Purification

Journal: Oncotarget

Article Title: Evaluation of a novel human IgG1 anti-claudin3 antibody that specifically recognizes its aberrantly localized antigen in ovarian cancer cells and that is suitable for selective drug delivery

doi:

Figure Lengend Snippet: OSPC-2 cells were incubated 1 h at 37°C with 2.5 μg/mL of IgGH6 in the presence of [10 μg/mL] FITC-CPE 290–319 peptide (green) panel A to allow antibody and peptide internalization, then cells were fixed, permeabilized and incubated with the Alexa594-conjugated secondary antibody (red) panel B. Nuclei were stained with DAPI (blue). Images were collected using a Zeiss Axiovert 200M epifluorescence microscope equipped with a Plan-Apochromat 63x/1.4 NA oil objective; z-stack images were acquired using ApoTome system and elaborated Inside4D module. panel C shows images resulting from merging the three different channels, with arrows showing IgGH6 and CPE peptide colocalization in the cell cytoplasm.

Article Snippet: After detachment with 0.5 mM EDTA, cells were incubated for 1 h with 2.5 μg/mL of purified IgGH6 followed by a 30 min incubation with mouse anti-human IgG-FITC secondary antibody (Southern Biotech).

Techniques: Incubation, Staining, Microscopy