ProtocolforAorticRingAssay
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) and excise thoracic aorta. Transfer the aorta into a dish with sterilized 1x PBS buffer. After removing the fibroadipose tissue, section arteries into 1-1.5 mm long cross sections, rinse the sections five times with endothelial cell growth medium (ECGM), and place the......閱讀全文
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
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
Adhesion－Assay－Protocol
Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini
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
alamarBlue?－Cell－Viability－Assay－Protocol
實驗概要Cell health can be ?monitored by numerous methods. Plasma membrane integrity, DNA ?synthesis, DNA content, enzyme activity, presence of ATP, and c
James－Hardwicks－angiotensin－assay－protocol
?This specific procedure was developed to assay the activity of the Lck kinase expressed from a retroviral vector in rat fibroblasts. You can obviousl
Roche公司的RNase－Protection－Assay－(RPA)－protocol
Roche公司的RNase Protection Assay (RPA) Using DIG-Labeled RNA Probes下載網址：http://www.roche-applied-science.com/PROD_INF/BIOCHEMI/no1_03/PDF/p22_23.pdf還有一份
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
Aspartate－Assay
實驗概要The ?Aspartate Assay Kit provides a simple, convenient assay to measure ?aspartate in a variety of samples. In the assay, aspartate is converted ?
Phosphate－Assay
1. Make standards using sodium phosphate at the following uM concentrations: 0, 2, 5, 7, 10, 20, 40, 60, and 80. Use the screw top glass tubes.2. Dry
Protease－assay
實驗概要? ? ? ? In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in
MTT－Assay
?This procedure is for cells in 96 well plates, if larger plates are used then adjust volumes accordingly.1 Make a solution of 5mg/ml MTT dissolved in
TUNEL－assay
PROTOCOL:?Deparaffinize and rehydrate slides:3 x 3′ Xylene3 x 2′ 100% ethanol1 x 2′ 95%, 80%, 70% ethanol (each)1 x 5′ 1x PBS?Microwave antigen retrie
Protease－assay
In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in helping to softe
Bradford－Assay
Bradford AssayThe bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie B
Polygalacturonase－assay
This enzyme is famous for being involved in the development of the GMO tomatoes (more information from the link at the foot of this page).?The cells o
Motility－Assay
DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these, acquisition o
Bradford－Assay
The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue
Pectinase－assay
Pectinases are actually a mixture of enzymes, which, along with others such as cellulase, are widely used in the fruit juice industry where they are w
DGK－Assay
Buffers:- 2X buffer10 ml 0.5 M imidazol, pH 6.60.21 g LiCl1.25 ml 1 M MgCl21.0 ml 0.1 M EGTA, pH 6.6--> Bring volume up to 50 ml with distilled water.
Chemotaxis－Assay
PurposeThe purpose of a chemotaxis assay is to determine whether your protein or small molecule of interest has chemotactic activity on a specific cel
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
關于RING－E3s的結構介紹
c—Cbl在與相應的E2發生作用時主要有兩部分發生作用，即酪氨酸激酶結合結構域(tyrosinekinasebindingdomain，TKB結構域)和RING結構域。TKB結構域有1個4H(four-helixbundle)，2個EF手形結構域和1個SH，結構域。RING結構域有1個3條鏈組成
概述RING－E3s的可能作用機制
RING家族E3一E2復合物的作用機制還不是很清楚，人們曾經認為RINGE3s通過別構激活泛素結合酶E2從而促進由E2直接催化的底物Lys殘基的泛素化，但在與c—Cbl相對應的泛素結合酶UbcH7結構上并沒有顯著的構象變化。RINGE3s的一功能是招募E2和底物，因此它可以通過增加E2周圍底物的
ELISPOT－Protocol
實驗概要The ?Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method ?of measuring the antibody or cytokine production of immune cells on t
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
ELISPOT－protocol
實驗概要The procedure ?below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits ?have been designed for detection of various cytokines and g
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
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.????????
ELISA－protocol
ELISA protocol：1.取5－10ul BMMY表達上清用0.05M NaHCO3稀釋到100ul鋪ELISA板，37度或室溫振蕩大于1小時。注意一定要做一個GS115空菌株表達上清作為陰性對照，最好還找一個帶有histag的蛋白作為陽性對照。2.TPBS洗板3次，方法：倒掉鋪板液，倒置于