ImmunohistochemistyFluorescenceProtocol1
MaterialsCytokine-specific Primary Antibodiesunlabeled or biotinylated antigen-affinity purified polyclonal antibodies (R&D Systems ''AF'' or ''BAF'' series) or selected monoclonal antibodies.Secondary Antibodies and Secondary ReagentsBiotinylated donkey anti-goat IgG (Jackson Immuno Research Lab Catalog # 705-066-147)Biotinylated goat anti-mouse IgG1 (Caltag Lab Catalog # M32115)F......閱讀全文
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
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
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
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 ?Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method ?of measuring the antibody or cytokine production of immune cells on t
Immunoprecipitation－Protocol
實驗概要Immunoprecipitation ?is a procedure by which proteins or peptides that react specifically ?with an antibody are removed from solution and examined
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
RNAi－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.????????
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
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
Cytotoxicity－Assays－Protocol
Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly, target cells were pelleted
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
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
Silver－Staining－Protocol
1x 40min - overnight?????50% MeOH, 12% Acetic Acid1x 30min??????????????????????????????????50% MeOH, 12% Acetic Acid, 0.05% 37% Formaldehyde3x 20min?
Transformation－Protocol－for－Arabidopsis
Transformation Protocol for Arabidopsis – AbbreviatedGerminate seed in pots↓ 4 weeksStreak bacteria onto YM/MinA↓ 2-3 days 28°CSpray/dip bacterial sus
Colony－PCR－Protocol
1. Pull out eight glycerol stock plates from the –80oC freezer and set on bench top to thaw. Be sure to remove the foil seal before leaving the plates
Immunofluorescence－Microscopy－Protocol
實驗概要Immunofluorescence ?allows the imaging of a specific factor in cells or tissue sections ?through the use of a specific antibody chemically which i
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
Nuclear－Extraction－Protocol
實驗概要The procedure presented below describes a method for extracting nuclear from several cell lines of human origin.主要試劑Hypotonic Buffer Solution20 mM
Immunofluorescence－Microscopy－Protocol
實驗概要Immunofluorescence ?allows the imaging of a specific factor in cells or tissue sections ?through the use of a specific antibody chemically which i
Western－Blotting－Protocol
實驗概要The western blot ?(sometimes called the protein immunoblot) is a widely used analytical ?technique used to detect specific proteins in the given s
Basic－ELISA－Protocol
實驗概要? ? ? ? There are many different types of ELISAs, which can detect the presence of protein in serum or supernatent. One of the most common typ
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
RNA－Isolation－Protocol
RNA Isolation Protocol(Revised 5-15-2003)Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109?cells (OD600= 0.2 Dilute cells or scale up)
Urea－Lysis－Protocol
Urea?lysis?buffer????????????9M Urea, 2.5mM EDTA, 2.5mM EGTA, 1% DTE, 4% CHAPS????????????make 10ml and aliquot 10x1ml, freeze at -70°C?Lysate?prepara
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
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
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
Yale－Immunofluorescence－Protocol
實驗概要We provide a protocol for fixation, immunostaining, and imaging in 384-well Plates.主要試劑Reagents1.?384-well view plates (Aurora)2.?HUVEC (pooled, L