免疫学技术

Analysis of total E. coli protein by SDS PAGEHahn Lablast modified 9/30/981. In microfuge tubes spin down 0.1 ml of uninduced cells grown to near saturation or 0.15 ml of IPTG induced cells. Remove YT ...

STAINING NONFILAMENTOUS ACTIN WITH VITAMIN-D BINDING PROTEINIndirect MethodMaterials1. Gc-globulin (vitamin-D binding protein; Calbiochem 345802) prepare 2 mg/ml stock in PBS solution A clarify 25000 ...

Vital Staining of Chromosomes with Hoechst 33258Materials1. Hoechst 33258 10 mg/ml stock in DMSO. 2. Culture medium. 3. 15 ml sterile conical tube. Procedure1. In a 15 ml conical tube add 1 volume Hoe ...

OUTLINEThe erythrophagocytosis assay is performed to compare the phagocytic rate with and without anti-eythrocyte antibody in either macrophages from control animals or virus-infected counterparts.PRO ...

OUTLINEClodronate (dichloromethylene diphosphonate) is used for treatment of osteolytic diseases and osteoporosis Its effect is due to inhibittion of the function of osteoclasts. This drug is also tak ...

Enrichment of PBMCs with monocytesContributed by Dr.A.GratchevThis protocol is used in our lab to reduce the costs of the cell sorting with MACS reagents. The cell suspension obtained after this proto ...

OUTLINEThis modification of peritoneal macrophage preparation is designed for a subsequent thrombophagocytosis test. PROTOCOLSacrifice a mouse (use dry ice CO2) Sink a mouse in the 5% water solution ...

Phagocytosis Fluorescent labeling of yeastResuspend 5 grams of yeast in 50 ml PBS in 100 ml flask Put flask for 30 minutes in boiling water bath while stirring. Wash 5 x with PBS and 2 x in PB. Adjust ...

Purification of human mononuclear cells and neutrophilsby Chang-Duk Jun 01/19/2000 PurposeMaterials10ml 6% dextran + 7ml citrate/citric acid Dextran: T500 -- 6g+100ml PBS Citrate solution: 25g Na Citr ...

Agglutination AssaysREFERENCE: Lanyi B. and T. Bergan. Methods in Microbiology Vol 10: 93-168. BACTERIAL AGGLUTINATION: Bacterial agglutination is performed using either P. aeruginosa cells autoclaved ...

SMEAR PREPARATIONThe 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 onto the slide and to prevent the ...

Flagella and motility monotrichous flagella - the bacterial cell has a single flagella peritrichous flagella - the bacterial cell has several flagella which are located at various sites on the cell su ...

Maintenance of Probes in bacteria including Escherichia coliPlasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured a ...

A. Phenol extraction of DNA samplesPhenol extraction is a common technique used to purify a DNA sample (1). Typically an equal volume of TE-saturated phenol is added to an aqueous DNA sample in a micr ...

Jeff Lawrence (M02)1. Place 2 ml of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed place up to 5 ml in a 16 mm green-capped culture tube. Large amount ...

Lyophilizing Cells Inoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized. Incubate the culture at 37°C with vigorous shaking (200-250 rpm) overnight. A ...

Jeff Lawrence (M05)1. Sterilize Velvet Squares with widths at least 50% longer than the diameter of a petri plate.2. Streak strains to be printed for single colonies.3. Prepare a printing Master Plate ...

Materials: Sterile LB medium (Luria-Bertani Medium) with or without antibiotic: water 500 ml bacto-tryptone 10 g bacto-yeast extracts 5 g sodium chloride 10 g stir to dissolve pH to 7.0 autoclave to s ...

Jeff Lawrence (M01)1. - From solid media touch an isolated colonies with a sterile applicator or toothpick.- From liquid media touch the culture with the end of a sterile applicator.- From a frozen cu ...

Agar Plates for Selection of Clones in BacteriaCloning of PCR productsStocks:LB Agar: Luria Broth after Lennox:per LiterTryptone10 gYeast Extract5 gSodium Chloride5 gBact. Agar15 gpH 7.0Autoclave ...