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Analysis of murine BM: specimen choice, sampling and processing.

Histologic analysis of murine BM is a necessary complement to flow cytometric or in vitro analysis. Techniques to do this are well established in human hematopathology. A long tradition in experimenta ...

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Pathology and Autopsy of a mouse

if mouse is found dead if mouse is a newborn or If mouse is alive or moribund and above one week of age Routine dissection Formalin fixation Storage of smears and slides Tissue embedding in paraffin ...

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Guidelines for Aseptic Rodent Survival Surgery

Introduction:Aseptic surgery is surgery performed without contamination or exposure to pathogens. These policies and guidelines are provided to help ensure optimal results for aseptic survival surgery ...

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Isolation of bone marrow

Isolation of bone marrow (contributed by Chris Jackson (chris.jackson@bris.ac.uk)) A protocol I have used to isolate rat bone marrow is: 1. Kill the rat and dissect out the femurs cutting the bones cl ...

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Isolation of colonic epithelium

The method we use is based on work of Dr. Hazel Cheng at the University of Toronto and works for both colon and the small intestine.First we excise the whole colon from cecum to rectum. This is easily ...

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Avertin (2, 2, 2-Tribromoethanol)

This is prepared as a 1.2% solution and used in mice at a dosage rate of 0.2 ml/10grams body weight which approximates 240 mg/kg. This compound is subject to photodegradation and should be stored at 4 ...

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Mouse Spleenectomy

PROTOCOLGeneral anesthesiainject i.p. 250µl/mouse of the ANESTHETIC mixture for verify the anesthetic effect check plantar reflexes (animal must be unresponsive to squeezing hind feet) Preparation of ...

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Guidelines for the Use of Anesthetics, Analgesics and Tranquilizers in Laboratory Animals

What is Anesthesia?Anesthesia is a state of unconsciousness induced in an animal. The three components of anesthesia are analgesia (pain relief) amnesia (loss of memory) and immobilization. The drugs ...

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Xenograft Tumor Model Protocol

Preparation of tumor cells Grow cells in complete medium and exclude any contamination When cells are 70-80% confluent 3-4 hrs before harvesting replace medium with fresh medium to remove dead and det ...

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Xenograft Tumor Assay Protocol

1) Determine the number of cells for injection (ie 5´106 ) to determine the number of plates thatwill require trypsinizing (usually a 100% confluent plate of 100mm2 will yield at least 2injections at ...

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Isolating Xenograft Tumor Cells for Tissue Culture or Transplantation

Isolation of T4 cells from xenograft tumorsPut mouse down Wash skin by dipping mice into ETOH Remove tumor and put into 5ml PBS containing Fungizone and Penstrep Swirl around to rinse tumor and pour ...

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Protocols for the preparation of tumour cells for s.c. injections in mice

For the establishment of solid tumour in nude mice tumour cells are treated in a sterile environment under a cell culturing hood. Disposable sterile plastic pipettes and flasks are used. Cultures are ...

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Basic Protein Chemistry Techniques

Coomassie Blue Stain: (for gels) 1) Combine 225 ml Methanol with 225 ml ddH2O. 2) Add 0.5 grams of Coomassie Blue. 3) Just before use add 50 ml acetic acid to 10% final concentration. 4) Stain for 2 ...

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Metabolic Labeling of Cells with 35S

1) Transfer to a 24 wells plate the desired colonies.2) Once the cells are attached (at least 8 hours after tripsinizing them) add ~1 ml ofDME (met- cys-).3) Add 3l of Met 35S Cys 35S trans label.4) I ...

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Serum Thawing Heat Inactivation

How to thaw serum:Serum that is stored at -10º C to -40º C is stable for extended periods of time. It is neither necessary or desirable to store serum at -70º C as it does not prolong the shelf life o ...

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Cell Culture Media and Solutions

Antimycotic/antibiotic media: To 1 liter of sterile RPMI 1640 with 2mM L-glutamine add: 165.0 ml fetal bovine serum heat inactivated 12.0 ml 200mM (100X) L-glutamine 12.0 ml antimycotic/antibiotic (10 ...

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TISSUE CULTURE STOCK SOLUTIONS AND MEDIA

MS MEDIUM FOR ARABIDOPSISTo 990 ml H2O add:Sucrose ........... 10.0 gMOPS .............. 0.5 gAgar .............. 8.0 gAdjust pH to 5.7 with 1 M KOH.Autoclave.When cooled add sterile stock soluti ...

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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 frozen rat liver PBS Homogenization Buffer Homogeniz ...

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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 0.02M Hepes and 1mM EDTA. pH=7.4 C. PBS with ...

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Large Scale Tubulin Preparation

Tubulin is purified from bovine/porcine brain by two cycles of polymerization/depolymerization followed by removal of copurifying proteins on a phosphocellulose (PC) column. The procedure described he ...

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