Journal: Scientific Reports

Article Title: Rigorous characterization of urinary extracellular vesicles (uEVs) in the low centrifugation pellet - a neglected source for uEVs

doi: 10.1038/s41598-020-60619-w

Figure Lengend Snippet: SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot (WB) analysis of P21 pellet. The whole pellet - originated from different volumes - was loaded in each lane and stained with either silver nitrate ( A ) or colloidal Coomassie ( B , C ). Nitrocellulose membranes were hybridized respectively with: collectrin (TMEM27) ( D ) and tissue inhibitor of metalloproteinases 2 (TIMP-2) and CD9. After the first image acquisition the same membranes were incubated again with insulin-like growth factor binding protein 7 (IGFBP-7) ( G ), podocin (NHPS2) ( H ) and calreticulin (CALR) ( I ) respectively. Finally membranes were incubated a 3 rd time with tumor susceptibility gene 101 (TSG101) ( J ), podocalyxin (PODXL) ( K ) and calnexin ( L ) respectively. Samples for gel in C and western blots in ( F , I , L ) were run without DTT. K rat kidney, S Saliva pellet 4.600 g.

Article Snippet: Nitrocellulose membranes were saturated with Odyssey blocking buffer (Li-Cor Biosciences) and incubated in 0.5 μg/mL rabbit anti podocin (NPHS2) (Code P0372), 1.0 μg/mL rabbit anti myosin IIA (MYH9) (Code M8064) and 0.5 μg/mL rabbit anti tumor susceptibility gene 101 protein (TSG101) (Code T5701) (Sigma-Aldrich,); 1.0 μg/mL rabbit anti insulin-like growth factor binding protein 7 (IGFBP7) (Code ab171085) and 1.0 μg/mL rabbit anti nephrin (NPHS1) [Y17-R] (Code ab136894) (Abcam, Cambridge, UK); 1.0 μg/mL goat anti metalloproteinase inhibitor 2 (TIMP2) (codeAF971) and mouse anti human serum albumin (ALB) (code MAB1455) (R & D System, Minneapolis, MN); 1.0 μg/mL rabbit anti collectrin (TMEM) in house [27]; 1.0 μg/mL mouse anti CD9 antigen (CD9) (code HBM-CD9-100) (HansaBioMed Life Sciences); 1.0 μg/mL mouse anti podocalyxin (PODXL) (code NBP2-33108), 1.0 μg/mL rabbit anti calreticulin (CALR NB600-101) and 1.0 μg/mL rabbit anti calnexin (CANX) (code NB100-1965) (Novus Biologicals) overnight at room temperature (RT) in the Odyssey blocking buffer diluted 1:1 with in house PBS (10 mM sodium phosphate dibasic, 1.8 mM potassium phosphate monobasic, 137 mM sodium chloride, 2.7 mM potassium chloride) and 0.15% (v/v) Tween-20.

Techniques: Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot, Staining, Incubation, Binding Assay

Journal: Journal of Thoracic Disease

Article Title: ERAS and patient reported outcomes in thoracic surgery: a review of current data

doi: 10.21037/jtd.2019.04.08

Figure Lengend Snippet: PRO definitions ( 17 )

Article Snippet: PRO Measure (PROM) , An instrument, scale, or single item measure used to assess the PRO concept as perceived by the patient, obtained by directly asking the patient (or in some cases a caregiver or surrogate) to self-report , PROMIS ® *.

Techniques: