PreparationofMouseNeutrophils
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 Island, NY] containing 0.1% bovine serum albumin and 5 mM HEPES.Percoll stock solution: the stock Percoll solution is made by adding 1 ml of 1.08 M NaCl to per 7 ml Percoll; store at 4 degree for less then 3 months.81, 62, 55, 50, and 45% Percoll solutions:......阅读全文
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－Mouse－Neutrophils
实验概要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 so
Purification－of－human－mononuclear－cells－and－neutrophils
PurposeMaterials10ml 6% dextran + 7ml citrate/citric acidDextran: T500 --> 6g+100ml PBSCitrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS4
Mouse－Spleenectomy
OUTLINEPROTOCOLGeneral anesthesia1. inject i.p. 250µl/mouse of the ANESTHETIC mixture < wait for 2-10min for the anesthetic effect>2. for verify the a
Pathology－and－Autopsy－of－a－mouse
if mouse is found deadif mouse is a newborn or < 1 wk of ageIf mouse is alive or moribund and above one week of ageRoutine dissectionFormalin fixation
Mouse－keratinocyte－cultures
PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al., Methods Enzymol
Isolation－of－mouse－embryos
1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forceps,use another to tear the mesome
Complete－Mouse－Necropsy
EuthanasiaEuthanasia and mouse necropsies require prior IACUC approval. The mode of euthanasia should be chosen which minimizes pain or distress to th
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
RNAse－A－Treatment－of－Mouse－Cells
IntroductionRNAse A treatment of permeabilized cells followed by immunostaining is a method which allows to show if the localization of a protein into
Mouse－Compete－Blood－Counts
Materials: 250 µL of fresh mouse blood in plastic tubes containing EDTA. RBC lysis buffer (388 mM NH4Cl, 29.7 mM NaHCO3, 25 µM Na2EDTA)20.75g. NH4CL2.
Chick－or－Mouse－embr...
实验概要The following procedure describes the procedure for whole mount staining of chick or mouse embryo’s. A similar procedure could be used for sta
Culturing－Mouse－Embryonic－Fibroblasts
MaterialsTrypsin (Gibco 25200-023)3T3 Medium: 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium + 20%
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－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