Invitrogen), protected from light. Then, cells were fixed with 2% paraformaldehyde for 15 min at room temperature (RT). Cells were permeabilized with 0.2% Triton X-100 in phosphate saline buffer (PBS
For visualisation of EGFP-HURP or endogenous HURP, cells were grown on acid-etched glass coverslips, permeabilized with 0.5 % Triton X-100 in 37 °C pre-warmed Microtubule Stabilising Buffer (MTSB buffer—80 mM KOH-PIPES (pH = 6.8), 1 mM MgCl2, 5 mM EGTA) for 30 s a...
使寡核苷酸对退 27 27 CN 116096904 A 说明书 13/52页 火,并随后使这些双链片段中的约5至6个通过黏性单链末端退火在一起,并随后将它们连 接在一起并克隆到标准细菌克隆载体中,例如克隆到可从Invitrogen Corporation, Carlsbad,Calif获得的 载体中。然后通过标准方法对构建体进行测序。制备由连 接在一起的80...
For transplantation experiments, the SVZs obtained from P2-5 WT or shootin1 KO mice were dissected and dissociated using trypsin-EDTA (Invitrogen) as previously described (Kaneko et al., 2010). The dissociated cells were labeled using the PKH-26 or PKH-67 Fluorescent Cell Linker Kit (Sigma)...
For microscopic imaging, slides were mounted with ProLong Gold Antifade reagent with DAPI (Invitrogen). Images were captured using a Leica TCS SP8 STED 3X confocal microscope with a 63x oil objective. G3BP1 Intracellular Phase Diagrams U2OS G3BP1/2 dKO cells were transfected with GFP-G3BP1 ...
DAPI Invitrogen D21490 Immunofluorescence DMEM gibco 41965-039 Cell culture DNAse Promega M6101 in vitro transcription DTT Promega included in P2077 in vitro transcription EGTA Roth 3054.3 Cytomix Escherichia coli JM109 Promega L2005 Transformation Fetal bovine serum gibco 10270106 Cell culture Fluoromoun...
an Antibody byMilliporeSigma (12G10)anti-alpha-tubulin an Antibody byDevelopmental Studies Hybridoma Bank (A-11008)Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 an Antibody byInvitrogen Antibodies (A-11011)Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibo...
Cell counting was performed using an automated cell counter TC20™ (Bio-Rad) loaded with a 1:1 ratio of Trypan blue staining (0.4%) (Invitrogen™, Thermo Scientific) plus cell solution on a counting slide. New passage number flasks were set up with 2 × 106 cells from the ...
powdered chow mixed with corn oil only. 5xFAD female mice aged 6 months were fed either with control AIN-76A standard chow or 0.12% PLX5622 (MedChemExpress, mixed in with AIN-76A standard chow, Research Diets) for 6 weeks. EdU (catalog no. E10187, Invitrogen) was dissolved in drinking...
HEK293T cells (Leibniz-Institut DSMZ) were cultivated at 37 °C and 5% CO2in DMEM Glutamax high glucose medium (Gibco), supplemented with 10% fetal bovine serum (Biowest). For transfection, cells were seeded and treated with Lipofectamine 2000 (Invitrogen) according to the manufacturer’s ...