harvest Protocols

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

There are two basic methods for the in vitro assay of B-galactosidase activity from yeast. They differ mainly in the method of preparing the material for assay. Both methods are described with accompanying protocols. Method I: Assay of Crude Extracts includes: Yeast Cell Growth; Yeast Cell Harvest; B-gal assays; Bradford Assays. Method II: Permeabilized cell assay. - [Read Assay of β-Galactosidase in Yeast Protocol]

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

Protocol for assay of β-Galactosidase in Yeast. Includes: Assay of Crude Extracts; Yeast Cell Growth; Yeast Cell Harvest; β-gal assays; Bradford Assays; Permeabilized cell assay. - [Read Assay of β-Galactosidase in Yeast Protocol]

Category: Cytogenetics Protocols

Cytogenetic protocol for blood preps. Includes: Preparation of materials; Culture Procedure - Use aseptic technique throughout procedure until harvest; Harvest; Slide making; Collecting blood by tail vein method; PHA Dose Response. - [Read Cytogenetic Protocol Blood Preps]

Category: C. elegans Protocols

Protocol for cytosolic extract & immunoprecipitation. Includes: Culture; Harvest; Extract; Immunoprecipitation. - [Read Cytosolic Extract & Immunoprecipitation Protocol]

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Immunoprecipitation Protocols

The use of denaturing Ips reduces background and simplifies protein harvest. Yeast are boiled in an SDS-containing solution to liberate total proteins. - [Read Denaturing Protein Immunoprecipitation from Yeast Protocol]

Category: Signalling Signal Transduction Protocols

This kinase assay is meant to determine whether an agonist or event can influence the autophosphorylation of FAK. The addition of 1 μl of polyGT to the kinase reaction mix will determine the activity of the enzyme against a substrate. Includes information on: Harvest, Immunoprecipitation, Kinase Reaction and Antibody Detection of FAK. - [Read FAK Autophosphorylation Assay]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, etc. Protocol provides a thorough description of LCM techniques, with an emphasis on tips and troubleshooting advice derived from LCM users. The total time required to carry out this protocol is typically 1–1.5 h. - [Read Laser-capture Microdissection Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: MEF Protocols Mouse Embryonic Fibroblasts

MEFs. Thompson Lab Stem Cell Methods and Procedure. Irradiation and plating MEFs, Splitting MEF cells, Thawing MEF, Freezing and Harvestion MEF cells. - [Read MEF Mouse Embryonic Fibroblast Protocols]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Following propagation to 1 X 108 cells, lymphoblastoid cells are conveniently stored at -80 degrees C to preserve the high molecular weight DNA in the cells until the DNA is purified. This procedure describes the steps required to harvest and freeze the c - [Read Method: Preparation of Lymphocyte Cell Pellet for Storage]

Category: Cytogenetics Protocols

Protocol for mitotic chromosome from cell cultures. Includes: Colchicine Treatment; Monolayer cultures; Cell Suspensions; Cell Harvest; Chromosome Slide Preparation. - [Read Mitotic Chromosome from Cell Cultures Protocol]

Category: Mouse Protocols: Mouse Cell Culture Protocols

Protocol for murine osteoblast harvest. - [Read Murine Osteoblast Harvest Protocol]

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

Protocol for osteoblast harvest (murine). - [Read Osteoblast Harvest (murine) Protocol]

Category: Cell Biology and Cell Culture: Hepatocyte Cell Culture

Process Protocol for Hepatocyte Harvest ... Re-suspend as necessary in culture media (for cell culture) or in CryoStor ... - [Read Process Protocol for Hepatocyte Harvest]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion. Includes: Antibodies for Lineage Depletion; Harvest Marrow; Lineage Depletion; Cell Sorting; Viral Transduction; Transplant; CFU assays; GFP and cell surface marker immunostaining. - [Read Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells for Culture, Flow Cytometry, or Transplantaio]

Category: Immunology

Protocol for purification of Lin-, c-kit+, Sca-1+ bone marrow cells for Culture, Flow Cytometry, or Transplantaion. Includes: Harvest Marrow; Lineage Depletion; Cell Sorting; Viral Transduction; Transplant; CFU assays; GFP and cell surface marker immunostaining. - [Read Purification of Lin-, c-kit+, Sca-1+ Bone Marrow Cells Protocol]

Category: Signalling Signal Transduction Protocols: MAPK Protocols

This protocol uses mitogen-activated protein/ERK kinase (MEK) to activate the extracellular-signal-regulated (Erk) mitogen-activated protein (MAP) kinases upon agonist binding to receptors. Protocol includes information about: Harvest, Immunoprecipitation and Kinase Reaction. Also includes following solutions: Laemmli Sample Buffer (1X), ATP Mix, Kinase Buffer (10X), Lysis Buffer. Helpful protocol hints are also included. - [Read Raf-1 Kinase Assay]

Category: Western Blot Protocols

Procedure describes how it denatures most of the modification and degradation proteins immediately giving the most accurate read out of the true levels of protein at the time of harvest.  However, in cases where detection is a problem, a limited purification (e.g. isolation of nuclear extract for the detection of transcription factors) might be required to allow analysis. - [Read Western Blot Analysis of Endogenous Gene Expression Protocol]