Eccles:ProteinLysatesfromCellsinCulture
Cell Lysis Buffer5mL 0.1M Tris HCl pH 8 (10mM)0.44g NaCl (150mM)0.02g EDTA (1mM)0.5mL nonidet P40 (1% w/v)0.05g SDS (0.1% w/v)Make up to 50mL with MQH2O and filter sterilise.Store at 4 degrees C.This cell lysis buffer has a limited shelf life (at 4 degrees C). I recommend making fresh buffer up annually. My own personal experience has been that after ~18months my protein lysates began appearing very distorted followi......阅读全文
Xenofree－Culture－of－Neural－Stem－Cells
实验概要Neural stem cells (NSCs) derived from human embryonic stem cells (hESCs) have the potential to help provide understanding for human neurogenesis
Primary－Culture－of－Pulmonary－Arterial－Smooth－Muscle－Cells
Primary Culture of Pulmonary Arterial Smooth Muscle Cells 1.Primary cultures of Pulmonary Arterial Smooth Muscle Cells (PASMCs) were isolated from hum
Isolation－and－culture－of－rat－coronary－microvascular－endothelial－cells
CMVE were isolated from Wistar rat hearts by digestion of primary cell isolation kit.1. Hearts mounted on a Langendorff apparatus were perfused at 3
Isolating－Xenograft－Tumor－Cells－for－Tissue－Culture－or－Transplantation
Isolation of T4 cells from xenograft tumors Put mouse downWash skin by dipping mice into ETOHRemove tumor and put into 5ml PBS containing Fungizone an
Isolation,－Culture,－and－Differentiation－of－Progenitor－Cells2
Slide Coating13. Just before use, dilute the poly-D-lysine stock solution 1:50 in H2O. Add 0.2 mL of diluted poly-D-lysine to each well of an eight-we
Cell－and－tissue－lysis－hub
This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison
Mesenchymal－progenitor－cells－derived－from－human－muscle
Harvesting traumatized muscle derived mesenchymal progenitor cells (MPCs)1. The muscle tissue was dissected without contaminating by granulation, adip
Dorsal－Root－Ganglion－Neuron－Culture－from－Adult－Rats
DescriptionProcedure for culturing and maintaining Dorsal Root Ganglion (DRG) neurons from adult rats. ProcedurePreparing Culture Surface- Collagen co
Identification－of－a－Mutant－Kinase/ATP－Analog－Pair2
Cellular Transfection and Immunoprecipitation Before proceeding with the experiments outlined below, all kinase pocket mutants should be characterize
Live－imaging－with－Drosophila－tissue－culture－cells2
Materials & ReagentsDrosophila Schneider S2 cellsSchneiders Medium (GIBCO/Invitrogen), 10% fetal calf serum, Antibiotics (Sigma A5955)Depression slide
Live－imaging－with－Drosophila－tissue－culture－cells1
IntroductionLive imaging provides an important complementation to the "snapshot" view obtained in fixed tissue by immunofluorescence. It allows follow
Isolation－and－culture－of－human－pulmonary－artery－smooth－muscle－cells－...
Isolation and culture of human pulmonary artery smooth muscle cells (PASMCs)1. The human pulmonary arteries were opened to expose the endothelial su
CO2恒温摇床解决人胚肾－293－(HEK293)－细胞结团问题（一）
人胚肾 293 (HEK293) 细胞在重组蛋白表达中是最常见的宿主细胞。 这类细胞能够表达大量的膜蛋白，如 G 蛋白偶联受体 (GPCR) ，是无法在最常见的生物制药生产宿主，如：中国仓鼠卵巢 (CHO) 细胞中作表达。 HEK293 虽然是蛋白表达的极好宿主，然而 HEK293 细胞
Isolation－of－T4－cells－from－xenograft－tumors
1 Put mouse down 2 Wash skin by dipping mice into ETOH 3 Remove tumor and put into 5ml PBS containing Fungizone and Penstrep 4 Swirl around to rinse t
Isolation－of－human－multipotent－mesenchymal－stem－cells－from－second
Isolation of human multipotent mesenchymal stem cells from second‐trimester amniotic fluid Culture of MSC from amniotic fluid1. Twenty amniotic flui
Derivation－of－Dopaminergic－Neurons－(from－Human－Embryonic－Stem－Cells)
实验概要Directed differentiation of specific lineages has been a focal point in the field of human embryonic stem cell (hESC) research. Cell replacement
Isolation－of－stromal－vascular－cells－from－human－adipose－tissue
Stromal stem cells proliferate in vitro and may be differentiated along several lineages. The scarcity of stromal stem cells in tissues and the la
Direct－PCR－from－Whole－Yeast－Cells:－Zymolyase－Method
Direct PCR from Whole Yeast Cells: Zymolyase MethodContributor: Namjin ChungDate: June 18, 19961. An average-size yeast colony (0.5-2mm) or a cell pel
Global－Expression－Profiling－of－RNA－from－Laser－Microdissected－Cells－at－...
Global expression profiling of RNA isolated from laser microdissected cells allows one to profile a specific set of cells allowing for enhanced se
Protein－Expression－and－Purification－Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T
Lambda（噬菌体）DNA－Miniprep
David HarryInstitute of Forest GeneticsUSDA Forest ServicePacific Southwest Research StationAugust 26, 1993Background :There are many published method
A－Method－for－Structure3
CSP-Dependent Transformation AssayTo determine if synthetic peptides activated quorum sensing for induction of genetic competence, we used the comC de
Culture－of－human－renal－proximal－tubule－cells－and－renal－cortical－fibroblasts
1. The method for primary culture of human renal proximal tubule cells (PTC) and renal cortical fibroblasts (CF) is described in detail.2. Renal cor
Ex－Vivo－Human－Primary－Mesenchymal－Stem－Cells－(MSCs)－Culture
Human bone marrow contains mesenchymal progenitors (mesenchymal stem cells, MSCs). MSCs produce adventitial cells in the human bone marrow microenvir
Protocol－for－Isolating－DNA－from－Blood－with－Nucleated－Red－blood－Cells
实验概要DNA isolation from fish or avian blood sample can be difficult because it contains nucleated red blood cells. E.Z.N.A. NRBC Blood DNA Kit is de
Western－blotting样品准备－（一）
实验概要Preparation of lysis buffers, protease and phosphatase inhibitors, lysate from cell culture, lysate from tissues, protein concentration, samples
Jacobs:Protocol－Total－Protein－Isolation－Using－RIPA－Lysis－Buffer
MaterialsRIPA buffer (RIPA buffer enables the extraction of cytoplasmic, membrane and nuclear proteins and is compatible with many applications, inclu
Protein－arginine－methylation
Typical modification sites: RGG box or RXR sequence motifs R-arginine, G-glycine,X-any aminoacid.Enzymes catalysing protein arginine methylation: PRMT
Immunoprecipitation－Protocol
实验概要Immunoprecipitation is a procedure by which proteins or peptides that react specifically with an antibody are removed from solution and examined
Isolation－and－culture－of－SpragueDawley－rat－aortic－smooth－muscle－cells
The intact mature arterial media is composed of at least four phenotypically unique cell subpopulations that reside in distinct medial layers. Th