TcellActivationProtocol
IntroductionMature T cells recognize and respond to the antigen/MHC complex through their antigen-specific receptors (TCR). The most immediate consequence of TCR activation is the initiation of signaling pathways including induction of specific protein tyrosine kinases (PTKs), breakdown of phosphatidylinositol 4,5-biophosphate (PIP2), activation of protein kinase C (PKC) and elevation of intracellular calcium ion con......阅读全文
Role－of－arrestins－in－the－activation－and－targeting－of－MAP－kinases
The binding of ?-arrestins to agonist-occupied GPCRs triggers the assembly of a MAP kinase activation complex using ?-arrestin as a scaffold, with sub
ELISPOT－Protocol
实验概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on t
ELISPOT－Protocol
实验概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on t
PCR－protocol
PCR reactionProtocol for 50µl reaction - adjust amounts if necessary, for a 20µl reaction use the same volumes of primer and dNTP-mix, but adjust the
ELISPOT－protocol
实验概要The procedure below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits have been designed for detection of various cytokines and g
Immunoblot－Protocol
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
ELISA－protocol
ELISA protocol：1.取5－10ul BMMY表达上清用0.05M NaHCO3稀释到100ul铺ELISA板，37度或室温振荡大于1小时。注意一定要做一个GS115空菌株表达上清作为阴性对照，最好还找一个带有histag的蛋白作为阳性对照。2.TPBS洗板3次，方法：倒掉铺板液，倒置于
RNAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
RLGS－protocol
A. Preparation of DNA SolutionIn the case of rice, for example This method may be appllicable for many grass species and some other plants.
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
Cellfree－System－for－the－examination－of－apoptotic－activity
IntroductionIn our lab we use the term 'cell-free system' when we talk about the examination of apoptotic activity in cytoplasmic extracts. T
Role－of－MAL－in－RhoMediated－Activation－of－SRF
Serum response factor (SRF) is a transcription factor, which binds to a serum response element (SRE) associated with a variety of genes including (i)i
Activation－of－cAMPdependent－protein－kinase,－PKA
G-protein coupled receptors (GPCRs) are one of the largest gene families of signaling proteins. Residing in the plasma membrane with seven transmembra
Activation－of－Src－by－Proteintyrosine－phosphatase－alpha
Progression through the cell cycle is accompanied by activation of the proto-oncogene c-Src, a protein tyrosine kinase. Overexpression of Src leads to
Lck－and－Fyn－tyrosine－kinases－in－initiation－of－TCR－Activation
T cell activation is initiated by recognition of antigen by the T cell receptor (TCR) in the context of Class II MHC on an antigen-presenting cell. Th
Aspirin－Blocks－Signaling－Pathway－Involved－in－Platelet－Activation
Activation of the protease-activated GPCRs in platelets contributes to platelet activation in clotting. The protease-activated receptors PAR1 and PAR4
Activation－of－PKC－through－G－protein－coupled－receptor
G-protein coupled receptors (GPCRs) transduce a variety of signals from the extracellular environment across the plasma membrane. One of the common si
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
Tissue－Harvest－Protocol
TISSUES TO BE PROCURED(minimally and preferably within 5-8 hours after death):1. Brain2. Liver3. Muscle4. Skin5: Others as the specific case dictatesP
Phycoerythrin－conjugation－protocol
Phycoerythrin conjugation protocolDavid's method modified from references (2) and (3). I used this method to conjugate a mouse IgG2a monoclonal an
Microarray－Hybridization－Protocol
Introduction:One microarray set consists of 7 nylon membranes with 2.5 x 7.5 cm dimension. 2304 genes were spotted onto nylon membranes (Schleicher an
Western－Blotting－Protocol
实验概要The western blot (sometimes called the protein immunoblot) is a widely used analytical technique used to detect specific proteins in the given s
Nuclear－Extraction－Protocol
实验概要The procedure presented below describes a method for extracting nuclear from several cell lines of human origin.主要试剂Hypotonic Buffer Solution20 mM
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
Protocol－for－dsRNA－Synthesis
实验概要 We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends
Migration－Assay－Protocol
Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)P
RNA－Isolation－Protocol
Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)Pipet 30 ml of RNAProtect Bacteria Reagen
BrdU－Labeling－Protocol
实验概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic
Western－Blot－Protocol
一、提取抗原蛋白将提取RNA途中留存的样品，加入150μl 100%酒精充分混匀，静置5min(RT), 2000×g , 4℃离心5min, 吸取上清至新管中, 加入750μl异丙醇, 混匀, 静置10min(RT), 12000×g, 4℃离心10min, 弃上清, 加入1ml 0.3mol/L