PreparationofLowDensity,CollagenaseDigestedSplenocytes
1. Dilute 2 ml of 4000 U/ml Collagenase D as follows: 1 ml into 9 ml HBSS/Ca++/Mg++ (=400U/ml) and 1 ml into 39 ml HBSS/Ca++/Mg++ (=100U/ml). Put on ice.2. Place 5 ml of the 100U/ml solution in a 10 cm dish. Over a dry dish, inject spleens with 100U/ml solution. Hold spleen with forceps and push onto 22-G needle while slowly injecting 1 ml collagenase solution. Tear open the spleen with the needle and transfer to dis......阅读全文
Preparation－of－Low－Density,－CollagenaseDigested－Splenocytes
1. Dilute 2 ml of 4000 U/ml Collagenase D as follows: 1 ml into 9 ml HBSS/Ca++/Mg++ (=400U/ml) and 1 ml into 39 ml HBSS/Ca++/Mg++ (=100U/ml). Put on i
Isolation－of－DL(Low－Density－Lipoprotein)－From－Human－Plasma
This procedure involves use of human plasma, a potentially dangerous source of blood-borne disease. Wear long-cuffed gloves, and eye-protection.Day on
Lowdensity－lipoprotein－(LDL)－pathway－during－atherogenesis
Low-density lipoprotein (LDL) is a plasma lipoprotein particle whose lipid component includes cholesterol and triglycerides and is commonly referred t
Isolation－of－murine－splenocytes
OverviewIn order to study spleen cells (e.g. lymphocytes, granulocytes, other immune cells), it helps to make single-cell suspensions so that the cell
Th9－Polarization－of－Mouse－Splenocytes
实验概要Th9 cells are a subpopulation of T-helper cells, characterized to be involved in allergic diseases and resistance against intestinal nematodes.
Sucrose－Density－Gradient－Fractionation－of－Yeast－Membranes
Grow a 2 ml culture, 24 hr. at 30oC in selective mediaWhen culture is ready, use it to inoculate about 55 ml (50 ml plus 5 for O.D.600 readings) of se
USE－OF－THE－LIGHT－MICROSCOPE
USE OF THE LIGHT MICROSCOPEEach time the microscope is to be used it should be set up correctly to give a good image. Most often users forget to adjus
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
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
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
Liposome－Preparation
Liposome PreparationOBJECTIVE:Method for incorporating proteins into liposomes for liposome swelling assay or as an alternative antigen presentation m
Template－Preparation
Template PreparationThe quality of sequencing results is directlyrelated to the quality of the template. ABI recommends a minialkaline-lysis/PEG preci
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
Hybridization－of－High－Density－Arrayed－BAC－Nylon－Filter－Blots
Protocol for hybridization of high density arrayed Bacterial Artificial Chromosome nylon filter blots with 100 PCR isolated Unigene cDNA inserts, pool
细胞培养常规操作
常规操作(主要内容如下)· Aseptic Technique· Culture Vessels· Cell Counting· Primary Culture· Maintenance of Cell Line ·
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
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
Competent－Cell－Preparation
实验概要Competent cells are those that possess more easily altered cell walls that DNA can be passed through easily. These cells readily incorporate f
CELL－MEMBRANE－PREPARATION
I. Solutions: A. Ca and Mg free Phosphate Buffered Saline (PBS) solution, buffered with 0.02M Hepes. pH=7.4 B. Ca and Mg free PBS, buffered with
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.
Preparation－of－Agar－plates
Prepare media and add 1.5 agar before autoclaving it (15g per liter). After autoclavation, cool the media in a 55 degree waterbath. Do not allow the s
Plasma－and－Serum－Preparation
实验概要Serum is the liquid fraction of whole blood that is collected after the blood is allowed to clot. The clot is removed by centrifugation and the
Preparation－of－Mouse－Neutrophils
实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline solution [HBSS; Invitrogen, Grand Isla
Preparation－of－human－platelets
Preparation of human platelets 1. Human blood was taken from drug-free volunteers on the day of the experiment using acidic citrate dextrose
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
Preparation－of－Polyacrylamide－Gels
1. Prepare 20X TBE as:216 g Tris Base110 g Boric Acid80 mL 500 mM EDTA, pH 8.0700 mL ddH2OMix. Bring volume to 1 L. Autoclave.2. Prepare Acrylamide so
Metaphase－chromosome－preparation
Materials: RPMI 1640 medium fetal calf serum (FCS), 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best.-Nr. 295892) cell cuture flask
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－MICROINJECTION－PIPETTES
INJECTION AND HOLDING PIPETTESThe glass capillary tubing used should be thin walled, borosilicate glass without a fibre.e.g. Clark Electromedical Inst
HELPER－PHAGE－PREPARATION
HELPER PHAGE PREPARATION1. Grow an overnight of NM522 in NZCYM medium.2. Dilute overnight 1:100 and grow to an A600 = 0.3 (@2.5 x 108 cells/ml).3. Inf