ELISAprotocol
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，倒置于毛巾上轻轻拍打几次，加TPBS至满，重复。3.加封闭液（TPBS加1％脱脂奶粉）350ml/孔，室温下放置40分钟－1小时，TPBS洗3次。4.加封闭液稀释的histag一抗100ul/孔，ELISA板于室温振荡0.5－1小时，TPBS洗3次。很多公司都有histag的单抗，个人觉得Qiagen和Pharmacia单抗较好，Invitrogen和Labvision的一般般，一般1：1000－5000稀释，不同公司的抗体效价不同。5.加封闭液稀释的HRP标记的羊抗鼠二抗100ul/孔，ELISA板于室温振荡0.5－1小时，TPBS洗3次......阅读全文
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
Basic－ELISA－Protocol
实验概要 There are many different types of ELISAs, which can detect the presence of protein in serum or supernatent. One of the most common typ
Sandwich－ELISA－Protocol
实验概要The Sandwich ELISA measures the amount of antigen between two layers of antibodies (i.e. capture and detection antibody). The antigen to be mea
ELISA－Protocol－(General－Guidelines)
实验概要Sandwich enzyme-linked immunosorbent assays (ELISAs) involve attachment of a capture antibody to a solid phase support. Samples containing known
ELISA－Protocol－(General－Guidelines)
实验概要Sandwich enzyme-linked immunosorbent assays (ELISAs) involve attachment of a capture antibody to a solid phase support. Samples containing known
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
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
RNAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
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
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
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
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
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.
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
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
Yale－Immunofluorescence－Protocol
实验概要We provide a protocol for fixation, immunostaining, and imaging in 384-well Plates.主要试剂Reagents1. 384-well view plates (Aurora)2. HUVEC (pooled, L
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
Silver－Staining－Protocol
1x 40min - overnight 50% MeOH, 12% Acetic Acid1x 30min 50% MeOH, 12% Acetic Acid, 0.05% 37% Formaldehyde3x 20min
Immunofluorescence－Microscopy－Protocol
实验概要Immunofluorescence allows the imaging of a specific factor in cells or tissue sections through the use of a specific antibody chemically which i
Intracellular－Staining－Protocol
1. Fix cells- Add16% formaldehyde directly into culture medium to obtain a final concentration of 1.5% formaldehyde.2. Incubate in fixative for 10 min
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
Bacteria－Culture－Protocol
Bacteria Culture ProtocolBy 徐晓政1、TBS Medium Preparation:Prepare 1L of TBS medium contains:Tryptone 12gYeast extract 24gNaCl 5gSodium Succinate 5gGlyce
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
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
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
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)
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