免疫学技术

Immunostaining of Cells Adherent to CoverslipsProcedure:1) Immerse 18 mm2 glass coverslips in EtOH. In the tissue culture hood individually pull out and carefully flame to sterilize. Allow to cool the ...

Introduction to ImmunocytochemistryImmunocytochemistry is the name given to methods that use antibodies to detect the location of proteins within cells. The antibodies bind specifically to the protein ...

General remarks:Labeled sections have to be fixed to avoid loss of label during the enhancement procedure and thoroughly washed with bi-distilled water to remove ions (especially chloride ions) which ...

Immunofluorescence on ultrathin resin sectionsThis protocol is taken from the 1999 FEBS course manual and was contributed by Heinz Schwarz.General remarks:Ultrathin methacrylate-(e.g.Lowicryl HM20 K4M ...

Antibody cleanupIn order to decrease the amount of nonspecific staining it is often necessary to preabsorb primary and secondary antibodies to yeast cells lacking the antigen prior to use. A 1:1 mixtu ...

Immunocytochemistry: Common problems and some answers.Immunocytochemistry at the ultrastructural level (TEM)Results obtained from immunocytochemical labeling can often differ from what was expected. S ...

Labeling Muscle Actin with 5-iodoacetamidofluoresceinDay 0 and 1Materials1. 0.5 mM ATP 0.2 mM CaCl2 2 mM Tris-HCl pH 8.0 at 4oC 250 ml for day 0. 2. 100 mM KCl 100 mM boric acid pH 8.4 at 4oC 10 ml. 3 ...

Preparation of Colloidal Gold Conjugates.Colloidal gold has been used for centuries in the preparation of stained glass for windows and fine glassware. In recent years colloidal gold particles have b ...

A semi-permanent mounting medium for immunofluorescence microscopyMaterials6gm glycerol 2.4gm mowiol 6ml distilled water 12ml Tris buffer 0.2M at pH 8.5 MethodWeigh out the glycerol in a 50ml plastic ...

Double immunofluorescence staining for BCL-6 and else.Any type of tissue is suitable for this technique as long as the antigenicity for your antigen(s) is preserved. Use cytocentrifuged cells or froz ...

FLUORESCENT STAINING OF CELLSFluorescent Phalloidin StainingMaterials1. Fluorescent phalloidin in methanol. Phallacidin does not work as well. Dilute 10 ul 330 nM stock into 500 ul PBS for each large ...

Fluorochrome Absorption and Emission SpectraHere we present the absorption and emission spectra of the fluorochromes BD Biosciences Pharmingen conjugates to monoclonal antibodies and other proteins. F ...

Anti-HA ImmunocytologyThe multiple copies of the HA epitope present in the HAT tag can be detected by the mouse monoclonal antibodies 12CA5 (Boehringer) and 16B12 (MMS101R; BAbCO Richmond California) ...

Yeast IF with Methanol/Acetone DehydrationDavid AmbergNote this protocol is a modified version of the protocol from Mark Rose in the CSH Yeast Genetics Course Manual.This is the protocol to use with t ...

Making Boiled Donkey (or whatever) Serum (BDS) for Blocking You want a final working concentration of 5% Serum. Order the serum from the same company and organism from which you buy secondary antibod ...

Preparing a cell block from cell lines / cell suspensions.Cell lines or purified cell suspensions are a valuable tool to provide known positive controls provide homogeneous tissue in unlimited amount ...

Staining for Surface Receptors contributed by Patricia TsaoThis protocols allows the use of fluorescence microscopy to visualize epitope-tagged receptors expressed by stable adherent cell lines. In co ...

Staining for total receptors contributed by Patricia TsaoThis protocol will visualize both internalized receptors that were once on the plasma membrane and receptors in the biosynthetic pathway. This ...

Figure 12.4 Typical bacterial shapes Materials Colonies of bacteria from Exercise 12.2ToothpicksCrystal violetGram's iodine95% ethanolSafraninMicroscopes with oil immersion Procedure Before staining ...

Gram-staining ProcedureGram-staining is a four part procedure which uses certain dyes to make a bacterial cell stand out against against its background. The specimen should be mounted and fixed on a s ...