CoIPProtocol2

Protocol1 调制protein A sapharoseprotein A sapharose 5ml 溶于20ml含0.02%NaN3的PBS离心2000转2分,去上清,在沉淀加0.02%NaN3的PBS,离心后去上清,重复2次,最后沉淀加20ml含0.02%NaN3的PBS,冷藏保存用单抗做一抗时,把protein A sapharose先用抗鼠Ig兔血清处理(每50ulprotein A sapharose用1-5ul血清)。旋转1-2h,混匀后,再离心1-2s去上清后,加PBS,vortex混合,离心去上清,重复二次加含0.02%NaN3的PBS到原来体积,冷存。避免长期保存。2.抗原的检出以下操作全在冰面或4度进行去除细胞培养液,用冷PBS洗一次。加RIPA,溶解后,转移到eppentube中用vortex混匀。RIPA的量:6cm的培养皿加1 ml(蛋白质的量为500mg/ml),组织块用液氮粉碎,在缓冲剂中捣......阅读全文

Dot-Blot-Protocol

a. Label nitrocellulose membrane (using a pencil) to identify protein elution fractions.b . Pipette 2μl from each fraction onto the membrane, allow th

Nucleolar-Isolation-Protocol

We recommend that you first download and read this page as a PDF file. Using that as your guide, you can then follow the protocol below and view a Qui

cDNA/AFLP-Protocol

Preparation of Para-magnetic beads from Promega cat#Z5482:a) suspend magnetic particles in bottle - transfer 200 ul (200 ug) of beads per sampleof RNA

TAIL-PCR-Protocol

TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 react

Protocol-for-Cell-Fusion

Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, the

IP防水防尘测试方法

IP6X将样品于接头组装成一体放入防尘箱中,测试中应用滑石粉,测试时间:8 小时没有灰尘侵入IPX8将样品与接头组装成一体装入试验工装内,从外接气插口向实验工装内注入清水,水面高于试件顶部至少20mm,水面上方留有一定的空间,在进气插头上插入进气管,另在工装漏水观察胶管插头接100mm 短管,从进气

大鼠IP3(IP3)ELISA试剂盒使用说明

原理本实验采用双抗体夹心 ABC-ELISA法。用抗大鼠 IP-3(interferon-inducible protein 3) 单抗包被于酶标板上,标准品和样品中的 IP-3与单抗结合,加入生物素化的抗大鼠IP-3,形成免疫复合物连接在板上,辣根过氧化物酶标记的Streptavidin与

大鼠IP10(IP10)ELISA试剂盒使用说明

原理本实验采用双抗体夹心 ABC-ELISA法。用抗大鼠 IP-10(interferon-inducible protein 10) 单抗包被于酶标板上,标准品和样品中的 IP-10与单抗结合,加入生物素化的抗大鼠IP-10,形成免疫复合物连接在板上,辣根过氧化物酶标记的Streptavi

Arabidopsis-RNA-extraction-protocol

1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below).

Streamlined-DNA-Extraction-Protocol

This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure wi

Arabidopsis-RNA-extraction-protocol

1-2 g fresh material, freezer-dried, ground with 0.2g sand (if necessary), and then homogenized with 10ml RNA extraction buffer (see below). Spi

Protocol-for-Construction-of-BAC-Libraries

Protocol for Construction of BAC Libraries       The bacterial artificial chromosome cloning (BAC) system is emerging as the system of choice f

Actin-StainingActin-Staining-Protocol

实验概要Invitrogen  offers several fluorescent and biotinylated phalloidin and phallacidin  derivatives for labeling F-actin. These phallotoxins, isolated

Cajal-Body-Isolation-Protocol

Buffers and solutions(All solutions are supplemented with Complete Protease inhibitor tablet (Roche, Cat no: 1-873-580) at the final concentration of

T-cell-Activation-Protocol

IntroductionMature T cells recognize and respond to the antigen/MHC complex through their antigen-specific receptors (TCR). The most immediate consequ

Marcantonio-Lab-Protocol-Manual

Protein GelPreparing and Running a Protein Gel (7% Polyacrylamide) A. Preparation of Running Gel Solution 1) Add to a 50 ml cylinder:DD-H2O 26 ml30% a

Protocol-for-Aortic-Ring-Assay

ProceduresCover a 48-well plate with Matrigel (100μl/well) of and incubate for 30 min at 37°C, 5% CO2.Sacrifice the1-2 month old mice/rats (WT/mutant

Simplified-Arabidopsis-Transformation-Protocol

实验概要Our present protocol (Clough and Bent, 1998; modified from Bechtold et al. 1993) is extremely simple. We have found that the MS salts, hormone

Tissue-preparation-protocol-for-ChIP

实验概要This protocol  describes how chromatin is prepared from tissue, which can subsequently  be used for chromatin immunoprecipitation (ChIP). It is re

Double-immunofluorescence:-sequential-protocol

实验概要We provide a protocol for immunofluoresent double staining incubating the antibodies separately.实验步骤1. Blocking and sequential incubation    1) Fi

Direct/Indirect-Staining-Protocol

Use this Protocol for Directly or Indirectly staining cells for Flow Cytometric Analysis1) Dilute cells to 5x10^6 cells/mL2) Aliquot 100uL of cells pe

Protocol-for-Protein-Extraction-for-proteomics

Protocol for Protein Extraction10 % w/v TCA/ acetone/ 0.07 % v/v -MercaptoethanolPlant cells are rich in compounds that interfere with the 2DE separat

Immunofluorescence-/-Confocal-Microscopy-Protocol

实验概要Immunofluorescence  is a technique used for light microscopy with a fluorescence microscope  and is used primarily on biological samples. This tec

Immunohistochemistry-Protocol-for-Frozen-Sections

实验概要The  following is a general procedure guide for preparation and staining of  acetone-fixed frozen tissues using a purified, unconjugated primary  

Native-chromatin-immunoprecipitation-protocol

实验概要The method is a native chromatin immunoprecipitation protocol.主要试剂1. 10 x TBS 0.1 M Tris-HCl (pH 7.5) 1.5 M NaCl 30 mM CaCl2 20 mM MgCl2 50 mM Na

RTPCR-PROTOCOL

RT-PCR PROTOCOL材料与方法…………………………………………………………    1.材料 ………………………………………………………1.1 供试用组织(细胞)…………………………………1.2 主要仪器设备………………………………………1.3 主要试剂……………………………………………1.

ImmunohistochemistyEnzymatic-Protocol

OverviewR&D Systems provides monoclonal, polyclonal and biotinylated antibodies for immunohistochemical use. The following protocol has been developed

Xenograft-Tumor-Assay-Protocol

1) Determine the number of cells for injection (ie 5´106 ) to determine the number of plates thatwill require trypsinizing (usually a 100% confluent p

Immunofluorescence-/-Confocal-Microscopy-Protocol

实验概要Immunofluorescence  is a technique used for light microscopy with a fluorescence microscope  and is used primarily on biological samples. This tec

Xenograft-Tumor-Model-Protocol

Preparation of tumor cellsGrow cells in complete medium and exclude any contaminationWhen cells are 70-80% confluent, 3-4 hrs before harvesting, repla