lb 005 human clusterin protein r d systems  (R&D Systems)

Journal: Diabetologia

Article Title: Exercise as a non-pharmacological intervention to protect pancreatic beta cells in individuals with type 1 and type 2 diabetes

doi: 10.1007/s00125-022-05837-9

Figure Lengend Snippet: Recombinant clusterin protects human beta cells against thapsigargin-induced apoptosis. EndoC-βH1 cells were pretreated for 24 h with 0 (white bars), 1 (grey bars) or 100 ng/ml (purple bars) recombinant clusterin. Medium was then changed, and cells were exposed or not to clusterin and thapsigargin (THAP, 1 μmol/l). Apoptosis ( a ) and DP5 expression ( b ) were analysed after 48 h. Gene expression was corrected for the geometric mean of the reference genes ACTB and VAPA and normalised to the condition THAP without clusterin set as 1. Results are mean ± SEM of six independent experiments. ** p <0.01 and *** p <0.001 compared with no clusterin/non-treated (NT); †† p <0.01 and ††† p <0.001 compared with no clusterin/THAP (ANOVA)

Article Snippet: EndoC-βH1 cells were pre-cultured for 24 h in medium supplemented with 2% FBS plus recombinant clusterin (no. 2937-HS-050, R&D Systems, Minneapolis, MN, USA) at concentrations of 1 and 100 ng/ml.

Techniques: Recombinant, Expressing, Gene Expression

Journal: Acta Neuropathologica Communications

Article Title: Clusterin ameliorates tau pathology in vivo by inhibiting fibril formation

doi: 10.1186/s40478-020-01079-1

Figure Lengend Snippet: CLU binds to tau and prevents tau aggregation. a In vitro tau fibrilization assay was performed with the 4R0N tau isoform in the presence or absence of recombinant human CLU and thioflavine T intensity was monitored during 24-h time course. b Quantification of thioflavine T signal at the 24-h time point. Four replicates were used. Data present as mean ± S.E.M. and analyzed by Student’s t test *** p < 0.001. c Western blot analysis of soluble (supernatant, s) and insoluble (pellet, p) fractions following tau fibril assembly in the presence or absence of exogenous CLU. d Tau assembly assay performed with 4R0N recombinant tau, tau PHFs from AD brains, and increasing concentrations of recombinant CLU. Four replicates were used. Data present as mean ± S.E.M. and analyzed by one-way ANOVA, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Tau filament assembly was performed with 5 μM 4R0N isoform in assembly buffer (10 mMHEPES (pH7.4), 33 mM NaCl, 1 mM MgCl2, 1.5 mM EGTA, 60 μM EDTA, 5uM Thioflavin-T and 20ug/ml Heparin) with and without 4 μM recombinant CLU (R&D #2937-HS-050).

Techniques: In Vitro, Recombinant, Western Blot