PreparationofLuciferinforInVitroandInVivoBioluminescentAssays
Preparation of Luciferin forIn Vitro Bioluminescent AssaysMaterials• D-Luciferin Firefly, potassium salt, 1.0 g /vial(Caliper Life Sciences Part Number #119222)• Sterile water• Complete mediaProcedure1. Prepare a 200X Luciferin stock solution (30 mg/ml) in sterile water. Mix gently by inversion until Luciferin is completely dissolved. Use immediately, or aliquot and freeze at -20 °C for future use.Note: One can eithe......阅读全文
In－Vitro－T－Cell－Activation
In Vitro T Cell ActivationIntroductionMature T cells recognize and respond to the antigen/MHC complex through their antigen-specific receptors (TCR).
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
DNA－Fragmentation－Assays－for－Apoptosis
Protocol I: Triton X-100 Lysis BufferIn 96 flat-wells plate, incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of eff
Coimmunoprecipitation－assays
co-IP assays can be performed between endogenous proteins or transiently or stably expressed exogenous - usually tagged - proteins. The advantage to u
SAPK/Jun－kinase－assays
Preparation of cell lysate:1. For cells in suspension, grow in DMEM with 10% FCS and then the day before you do the experiment spin the cells down (1
PLAQUE－ASSAYS－FOR－ADENOVIRUS－TITRATION
-Set up 60 mm dishes of P11 cells to be 100 confluent at time of infection. -Remove medium from dishes, add 0.2 to 0.5 ml virus and adsorb for 30 – 60
cAMP分析－cAMP－Assays
cAMP AssaysGouzel Karimova and Daniel LadantUnite Postulante de Biochimie des Interactions Macromoleculaires, Departement de Biologie Structurale et C
DNA－Immunoprecipitation－for－the－Determination－of－DNABinding－Specificity
Andrea J. Gossett and Jason D. Lieb1Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA1Corresponding autho
In－Vivo－Imaging－of－Far1
In Vivo Imaging of Far-red Fluorescent Proteins after DNA Electrotransfer to Muscle TissueDNA electrotransfer to muscle tissue yields long-term, high
In－Vivo－Imaging－of－Far3
To determine the minimum dose of Katushka plasmid needed to give detectable fluorescent intensity, we decreased the amount of pTurboFP635 to 0.5 and 1
In－Vivo－Imaging－of－Far2
In vivo bio-imaging Mice were anesthetized and placed in a custom-made bed, which allowed stable and reproducible imaging of the legs. In vivo scann
Preparation－of－nucleic－acid－probes
Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s
Preparation－of－tubulin
Although many protocols for tubulin preparation are available, the procedure described below is the simplest and highest yielding preparation I have d
CAM－preparation
8 eggs per day, day 7- day 13 cut CAM, wash in precooled PBS,in 10 ml WASH 1 (PBS, 5 mM EDTA, COMPLETE) on icecut in pieces in petri dish on icecentri
SMEAR－PREPARATION
The preparation of a smear is required for many laboratory procedures, including the Gram-stain. The purpose of making a smear is to fix the bacteria
Template－Preparation
Template PreparationThe quality of sequencing results is directlyrelated to the quality of the template. ABI recommends a minialkaline-lysis/PEG preci
Platelet－Preparation
OUTLINEIn order to avoid platelet activation all manipulations must be performed as quickly and as acurate as possible.Work on ice if possible! This p
Liposome－Preparation
Liposome PreparationOBJECTIVE:Method for incorporating proteins into liposomes for liposome swelling assay or as an alternative antigen presentation m
Invitro－Phagocytosis－Assay－of－Macrophages
IntroductionThe term phagocytosis itself describes its mean phage = engulfment; cytosis: cell process. In other words, phagocytosis is the cellular pr
In－Vitro－Conservation－and－Cryopreservation－of－Ornamental－Plants
Today, the conservation of ornamental germplasm can take advantage of innovative techniques which allow preservation in vitro (slow growth storage
Microtubule－(MT)/Organelle－Motility－Assays
Rapidly thaw and immediately place on ice one aliquot each of axonemes, Golgi or ER membranes, 45 uM tubulin, rat liver cytosol, and 20x energy regene
Nuclear－RunOn－Transcription－Assays
Nuclear “run-on” (or “run-off”) transcription assays have been used to obtain quantitative information about the relative rates of transcription o
ex－vivo－expanded－endothelial－progenitor－cells
Cell Culture. 1. Total hPBMCs were isolated from blood of human volunteers by density gradient centrifugation. 2. Cells were plated on culture dishes
体外转录
· In Vitro RNA Transcription (Promega)For in vitro preparation single-stranded RNA probes or microgram quantities of defined RNA transcripts f
PREPARATION－OF－MICROINJECTION－PIPETTES
INJECTION AND HOLDING PIPETTESThe glass capillary tubing used should be thin walled, borosilicate glass without a fibre.e.g. Clark Electromedical Inst
DGK－Membrane－Preparation
Reagents:Bacterial strainE. coli N4830/pJW10LB amp media50 µg/ml ampicillinHigh salt bufferfor 1 L50 mM KH2PO4 6.8 g150 mM KCl 11.18 g50 mM sodium pyr
Preparation－of－Phage－Lysates
Preparation of Phage LysatesInoculate 5 ml of lambda-broth in a glass culture tube with a single colony of an appropriate host strain of E. coli. Incu
Lambda－DNA－Preparation
Lambda DNA PreparationThis is a plate method that gives very good yield for cloning. I have combined the Promega and Maniatis protocols.Solutions T-TY
PREPARATION－OF－SEQUENCING－GELS
MATERIALS:2-glass plates1 sharks -tooth comb and spacersWhatman 3 mm paper30 or 40% acrylamide-bis (19:1)10X TBEurea10% ammonium persulfateTEMED60 cc.
Rat－Liver－Preparation
实验概要The procedure presented below describes a method for preparing rat liver.主要试剂1. Aluminum Foil2. Liquid Nitrogen3. Dry Ice4. Ph