细胞技术

Steven Finkbeiner Departments of Neurology and Physiology UCSF Protocol note: hood scgedule for day of transfection: - 6h gap - CELLS: Polylysine coat plates. add 5ml polylysine per 100mm plat ...

Principle: Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from anticoagulated venous blood are isolated by layering onto histopaque. During centrifugation eryth ...

Chen-Ming Fan LabCarnegie Institute of Washington Prepare cell lysate as you would for Luciferase assay. Take 50uL of lysate and dilute with 100uL of 1X Reporter Lysis buffer (brown bottle at room ...

Seed cells at appropriate density (5x104/well for 3T3 MPAC BHK; 5x105/well for aTC1.6 bTC3) one to two days before transfection. Combine total DNA at 1ug /well with 600ul/well serum-free medium in an ...

Transfect 5 ug of the desired plasmid to the desired adherent cell line. Transfect in parallel GFP and carrier DNA (total of 5 ug). 48hrs after transfection check for transfection efficiency. Takin ...

一、基本概念 (一)基因导入(或基因转导) 将外源性基因(或基因组DNA)采用分子生物学技术人工地导入细胞，观察它在细胞中的表达，研究其生物学特性和功能，这种把基因导入细胞中的技术称为基因转导(gent transfer)或称基因转移，也简称为导入。是研究基因表达、结构和功能的重要研究手段。 (二)基因转导的种类： 目前基因转导技术很多，可根 ...

Introduction Quality is important in all aspects of tissue culture since the quality of materials used i.e. media and other reagents) will affect the quality of the cultures and products derived from ...

1. Dilute 2 ml of 4000 U/ml Collagenase D as follows: 1 ml into 9 ml HBSS/Ca2+/Mg2+ (=400U/ml) and 1 ml into 39 ml HBSS/Ca2+/Mg2+ (=100U/ml). Put on ice. 2. Place 5 ml of the 100U/ml solution in a 10 ...

Material Instruments Solutions 1 Curve Scissor ...

Procedure 1. Pour buffy coat (see Hint #1) from the bag into a 125 ml conical polypropylene centrifuge tube and dilute to 90 ml with sterile endotoxin-free PBS. 2. Add 15 ml of sterile Dextran/PBS an ...

METHOD: 1.Established hybridomas and other cell lines can be stored in liquid N2 using this method. 2.Have everything ready before you start working with the cells (i.e. cool media label vials prepare ...

Freezing Cells: 1.Trypsinize cells and harvest in the normal way. 2.Count a 200 μl aliquot and determine the total cell number.From thiscalculate the volume of media required to give a final freezi ...

The kitchen makes Tris TD Tryp/TD PBS and VE. Tris is a fairly complex Tris-buffered physiological saline solution. It is used to wash cells and is preferable to TD unless you need a Ca2+/Mg2+ free s ...

MS MEDIUM FOR ARABIDOPSIS To 990 ml H2O add: Sucrose ........... 10.0 g MOPS .............. 0.5 g Agar .............. 8.0 g Adju ...

最近看大家常提到支原体污染，就翻了翻书，做了些笔记，现在贴出来和大家分享： 支原体是一种大小介于细菌和病毒之间(最小直径0．2um)并独立生活的微生物．约有1%可通过滤菌器。支原体无细胞壁形态呈高度多形性．可为圆形、丝状或梨形。 支原体形态多变，在光境下不易看清内部结构。电镜下观察支原体膜为三层结构．其中央有电干密度大的密集颗粒群或丝状的中心束。支原体多吸附或散在于细胞表面和细胞之间。横断面与细 ...

Role of Rho Rac and Rho-kinase in phosphorylation of myosin light chain development of polarity and spontaneous migration of Walker 256 carcinosarcoma cells. As previously shown constitutive activatio ...

关于293/293T细胞 1、种属都为人，不是鼠。 2、来源：293细胞是转染腺病毒E1A基因的人肾上皮细胞系，293T细胞由293细胞派生，同时表达SV40大T抗原，含有SV40复制起始点与启动子区的质粒可以复制。用Ca3(PO4)2转染效率可高达50%。蛋白表达水平高，转染后2-3天用碱性磷酸酶分析可较容易地检测到表达的蛋白。瞬时转染293T细胞是过表达蛋白并获得细胞内及细胞外（分泌的或膜）蛋 ...

CHO-K1 (hamster Chinese ovary) ECACC 85051005 Morphology: Epithelial Hamster Chinese ovary Depositor: Dr D Newell PHLS CAMR Porton Down Salisbury UK No restrictions. Patent: None Specified By Deposito ...

This protocol is for use with the D cyclins and employs 488 nm argon laser excitation of propidium iodide and 630 nm NeNe or diode laser excitation of the fluorochrome Cy5 to detect cell cycle-specifi ...

1.Add 100 l of well-mixed anticoagulated whole blood to the bottom of a labeled tube. 2.Add the appropriate primary antibody to each tube. If using unlabeled antibody a titration is suggested. Conjug ...