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细胞功能测定

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Tissue Culture Methods

TYPES OF CELLS GROWN IN CULTURETissue culture is often a generic term that refers to both organ culture and cell culture and the terms are often used interchangeably. Cell cultures are derived from ei ...

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Preparation of Feeder Layers And SNL Stocks

Preparation of Feeder Layers And SNL Stocks1.1. Gelatinizing PlatesPrepare plates by covering surface with 0.1% Gelatin solution: Plate Size (cm) Amount of Gelatin (ml) 3 2 6 4 10 9 15 18Swirl the pla ...

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Preparation of NeoR Murine Embryonic Fibroblasts

Preparation of NeoR Murine Embryonic FibroblastsThree weeks prior to embryonic fibroblast isolation a PGK-neo male mouse is mated to a heterozygote female. 13.5 days to 14.5 days after the plug is obs ...

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Differentiate ES cells into glial cells and neurons

Differentiate ES cells into glial cells and neuronsDay -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.___________________Day 1: Trypsin ...

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Isolation of Primary Fibroblasts from Mouse Embryos

Isolation of Primary Fibroblasts from Mouse Embryos 1. Treat 10 cm tissue culture plates with 0.1% gelatin for at least two hours before use. 2. Sacrifice the pregnant female mouse (day 13 or 14 p.c.) ...

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PRODUCTION OF ES CELL CHIMERAS BY AGGREGATION WITH EIGHT-CELL STAGE EMBRYOS

Introduction The technique described here is a slight modification (March 1997) of methods presented in: Nagy A. J. Rossant. 1993. Production of completely ES cell-derived fetuses. InGene targeting ...

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Immunofluorescent Staining of Mouse and Rat Leukocytes

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 sus ...

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Phenotype-specific immunodetection of cyclins using 488/630 nm dual laser flow cytometry

Phenotype-specific immunodetection of cyclins using 488/630 nm dual laser flow cytometry William Telford Hospital for Special Surgery This protocol is for use with the D and E cyclins and employs 488 ...

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Conjugation of monoclonal antibodies

Conjugation of monoclonal antibodiesPrelude: You are free to copy and distribute these documents at will--but please do so in their entirety complete with figures. To reference this material please in ...

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Simultaneous analysis of DNA content and surface

Simultaneous analysis of DNA content and surface immunophenotype using gentle ethanol fixation techniques. William Telford. Louis E. King and Pamela J. Fraker Michian State University East Lansing M ...

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Staining Procedure for Flow Cytometric Detection of Human Cyclins

Staining Procedure for Flow Cytometric Detection of Human CyclinsThis is a standard protocol used at Pharmingen for Quality Control testing of the anti-cyclin antibodies by flow cytometry. It is a goo ...

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FACS Analysis Using Peripheral Blood Cells

FACS Analysis Using Peripheral Blood CellsCollect blood (75 microliters) into 1ml PBS containing 5 microM EDTA (10 microliters of 0.5 M stock) and mix immediately to prevent clotting. Keep tubes on ic ...

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Timing of Cycles

Materials Monolayer cultures grown in 75 mm culture flasks (Cells from Exercise 11.4 may be used or cultures of tetrahymena yeast or algae may be used.)H-thymidine with at least 4 µc/ml 0.36 c/mMPhosp ...

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Tissue Culture - Thawing Cells from Liquid Nitrogen

Tissue Culture - Thawing Cells from Liquid NitrogenMETHOD:Prepare before thawing:Fill a test tube of 10 - 15 mls of cold media (appropriate for your cell line) with 10 - 20% fetal calf serum (use 2x t ...

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Immuno-Laser Capture Microdissection

A: Development of Immuno-LCM Limitation of Microdissection Microdissection of routinely stained or unstained frozen sections has been used successfully to obtain purified cell populations for the anal ...

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Laser Capture Microdissection (LCM)

This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor the following protocol for their own research obj ...

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Guide to Cell Proliferation and Apoptosis Methods

Chapter 1: Cell Death - Apoptosis and Necrosis 1.1 Introduction21.1.1Terminology of cell death 21.1.2Differences between necrosis and apoptosis31.1.3Apoptotic Pathways4 1.2 Apoptosis Assay Methods6 Me ...

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Detection by TUNEL labeling

In Situ Cell Death (Apoptosis) Detection by TUNEL labelingby Boehringer Mannheim (Catalog No. 1684809) modified by Josiah N. WilcoxJosé C. Rodriguez and Héctor de Léon - Emory University - April 1996P ...

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TUNEL labeling

In Situ Cell Death (Apoptosis) Detection by TUNEL labelingby Boehringer Mannheim (Catalog No. 1684809) modified by Josiah N. Wilcox andJosé C. Rodriguez - Emory University - April 1996Protocol for Fro ...

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Staining Methods for cell

death Z. Xia 10/2/95The simplest way: trypan blue. Dead cells stain blue Non-fixed cells: FDA(fluorescein diacetate)-green alive cells; P.I. (propidium iodide)-red dead cells 35 mm plates: to 2 ml med ...

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