ImmunofluorescentStainingofMouseandRatLeukocytes
I. ProcedureHarvest cells from tissue, preparing a single cell suspension. Red blood cells may be removed by lysis or density gradient: Red blood cells from murine peripheral blood or a spleen cell suspension can be lysed using BD Biosciences Pharmingen's PharM Lyse™ (Cat. No. 35221E) solution. Add 2.0 ml of 1X Lysing Solution to the spleen cell suspension or per 200 µl of murine peripheral blood. Gently vortex i......阅读全文
Actin－StainingActin－Staining－Protocol
实验概要Invitrogen offers several fluorescent and biotinylated phalloidin and phallacidin derivatives for labeling F-actin. These phallotoxins, isolated
Gramstaining－Procedure
Gram-staining is a four part procedure which uses certain dyes to make a bacterial cell stand out against against its background. The specimen should
Intracellular－Cytokine－Staining－Protocol
实验概要A modification of the basic immunofluorescence staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surf
Direct/Indirect－Staining－Protocol
Use this Protocol for Directly or Indirectly staining cells for Flow Cytometric Analysis1) Dilute cells to 5x10^6 cells/mL2) Aliquot 100uL of cells pe
High－resolution－negative－staining
High resolution negative staining(From Valentine et al, 1968. Biochemistry 7:2143-52)Rationale: For the highest resolution with negative staining, the
Blood－Smear:－Preparation－and－Staining
Blood Smear: Preparation and StainingReference:Davidson, I. and Henry J., Clinical Diagnosis by Laboratory Methods, I. Davidsohn and J. Henry, eds., W
Preparation－and－Staining－of－Paraffin－Sections
I. Fixation and Processing of Tissue for Paraffin SectionsA. Fixation of Tissues in 10% Neutral Buffered FormalinSacrifice animal by prescribed and ap
Staining－Methods－for－cell－death
The simplest way: trypan blue. Dead cells stain blueNon-fixed cells: FDA(fluorescein diacetate)-green, alive cells; P.I. (propidium iodide)-red, dead
Rat－Blood－Collection－Protocols
实验概要The procedure presented below describes a method for collecting rat blood.实验步骤Rat should be fully anesthetized (e.g., unresponsive to toe pinch).1
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
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%
Intracellular－Immunofluorescent－Staining－for－Flow－Cytometry
IntroductionA modification of the basic immunofluorescent staining and flow cytometric analysis protocol can be used for the simultaneous analysis of
常用的βactin－引物序列
human actin f ctc cat cct ggc ctc gct gt human actin r gct gtc acc ttc acc gtt cc product size:268 rabbit actin r agt gcg acg tgg aca tcc g rabbit act
常用的βactin－引物序列
human actin f ctc cat cct ggc ctc gct gt human actin r gct gtc acc ttc acc gtt cc product size:268 rabbit actin r agt gcg acg tgg aca tcc g rabbit act
Isolation,－Culture,－Characterization－of－Cortical－and－Hippocampal－Neurons
实验概要The ability to culture primary neurons under serum-free conditions facilitates tighter control of neuronal studies. Some serum-free media and s
Examination－of－a－Mammalian－Blood－Smear
Examination of a Mammalian Blood SmearThe distribution and appearance of the formed elements of blood can tell a great deal about the condition of the
Methylene－Blue－DNA－staining－protocol
Methylene Blue DNA staining protocolProtocol:Load 2-5X the amount of DNA that would give bands of moderate intensity on an ethidium bromide stained ge
Preparation－and－Staining－of－Frozen－Tissue－Sections
I. Preparation of Frozen Sections for SectioningMaterials neeed:2-methylbutane (isopentane)Liquid NitrogenDry icePeel-Away?/sup> base moldsFrozen tiss
SSR－GEL－and－Silver－Staining－Protocol
I. EQUIPMENT:DNA sequencing unit (35 x 45 cm) & 2000V power supplyClampsLg. plastic trays (4), about 43 x 50 x 8 cm, and one lidTwo rocking platformsH
Immunofluorescent－staining－of－Sea－Urchin－embryos
1. Transfer fixed embryos to microfuge tubes. Allow to settle for 10 minutes.Gently remove most of the liquid.2. Add 100 ul antibody to one tube and 1
Isolation－of－rat－cardiac－fibroblasts－and－cardiomyocytes
1. Hearts were removed from newborn rats (day 0), put into calcium- and bicarbonate-free HEPES-buffered Hanks’ medium, cut into pieces and digeste
Rat－urinary－bladder－urothelial－cells
1. Bladders were excised from deeply anesthetized (urethane, 1.2 gm • kg−1, i.p.) Sprague Dawley rats (of either sex), cut open, and gently stretc
Preparing－Mitochondria－from－Rat－Liver
Liver is a convenient source for functional intact mitochondria for a number of reasons. Animal tissue is more readily homogenized than plant tissue b
Preparation－of－Rat－Liver－Cell－Cytosol
These protocols should yield enough cytosol and organelles for 1-200 MT/Organelle motility assays.Solutions and Reagents Freshly removed or flash fro
Preparation－of－Mitochondria－from－Rat－Liver
Preparation of Mitochondria from Rat LiverRat liver is an ideal source for functional intact mitochondria for a number of reasons. We use Sprague-Dawl
Histopathological－Approach－to－Rat－Liver－Tissue
ProcedureAfter deep ether anesthesia, dissect the rat’s liver (Wistar albino rats, 200 – 250 g) by cutting on the ventral side.Fix 2 – 3 mm. of the li
Peripheral－blood－“endothelial－progenitor－cells”
EPC Isolation and Characterization1. EPCs were obtained by isolating mononuclear cells using Ficoll density-gradient centrifugation of human blood buf