sucrose Protocols

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Bradford Assay Method - http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Biochemistry/Bradford.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols: Bradford Protein Assay Protocols

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue to protein. There is no interference from cations nor from carbohydrates such as sucrose.
Detergents such as sodium dodecyl sulfate and triton x-100 can interfere with the assay, as well as strongly alkaline solutions.

Includes a general overview of the procedure and preparation of the standards in the protocol. - [Read Bradford Assay Method]

Cajal Body Isolation Protocol - http://www.lamondlab.com/f7cbprotocol.htm

Category: Cell Organelle Protocols: Nucleus Protocols

Cajal Body Isolation Protocol. Protocol includes: Sonication, Removal Nucleoi, Gradient One, Gradient two, Concentration and final enrichment of cajal bodies. Also includes: Making 2.55M sucrose stock and Analysis of the enriched Cajal body fraction. - [Read Cajal Body Isolation Protocol]

CHO Centrosome Prep Protocol - http://mitchison.med.harvard.edu/protocols/mt1.html

Category: Cell Organelle Protocols: Nucleus Protocols

The key step is the lysis which solubilizes centrosomes away from nuclei by very low ionic strength lysis after treatment of cells with nocodazole and cytochalasin B. The released centrosomes are then centrifuged onto a Ficoll cushion (to avoid pelleting) and the interface between the lysate and the Ficoll is collected and the centrosomes are concentrated on a sucrose gradient. Fractions are assayed by spindown and double IF with 5051 serum and anti-tubulin and the pooled fractions are frozen... - [Read CHO Centrosome Prep Protocol]

Cleaning Worms by Floating on Sucrose Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=18

Category: C. elegans Protocols: C. elegans Cell Culture Protocols

Protocol for cleaning worms by floating on sucrose. - [Read Cleaning Worms by Floating on Sucrose Protocol]

Cryosectioning Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/C7.html

Category: Immunohistochemistry Protocols: Cryosectioning Protocols

Protocol for cryosectioning. While the timing of the various steps in this protocol are probably not critical, process the tissue all at once to ensure that RNA and/or proteins do not get degraded. Includes: 20% Paraformaldehyde/4% Paraformaldehyde-PBS; Sucrose/PBS. - [Read Cryosectioning Protocol]

Drosophila Cornmeal, Sucrose and Yeast Medium Protocol - http://fly.bio.indiana.edu/Fly_Work/media-recipes/harvardfood.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila cornmeal, sucrose and yeast medium. - [Read Drosophila Cornmeal, Sucrose and Yeast Medium Protocol]

Drosophila Cornmeal, Sucrose, Dextrose, Yeast and 2-Acid Medium Protocol - http://fly.bio.indiana.edu/Fly_Work/media-recipes/caltechfood.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila cornmeal, sucrose, dextrose, yeast and 2-acid medium. - [Read Drosophila Cornmeal, Sucrose, Dextrose, Yeast and 2-Acid Medium Protocol]

Drosophila Sucrose and Yeast Medium Protocol - http://fly.bio.indiana.edu/Fly_Work/media-recipes/neurofood.htm

Category: Drosophila Protocols: Drosophila Culture Media Protocols

Protocol for Drosophila sucrose and yeast medium. - [Read Drosophila Sucrose and Yeast Medium Protocol]

Equilibrium Density Gradient Percoll Protocol - http://homepages.gac.edu/~cellab/chpts/chpt3/ex3-3.html

Category: Cell Biology and Cell Culture

Cell fractionation of cellular components using Percoll a synthetic, colloidal solution of polyvinylpyrrolidone coated silica, specifically designed for sedimentation centrifugation. Percoll becomes a simple matter to establish a linear density gradient. Organelle separations are much easier to accomplish on Percoll density gradients than on sucrose gradients. - [Read Equilibrium Density Gradient Percoll Protocol]

Extraction and Solubilization of Total Protein from Microorganisms Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4224

Category: Bacterial Cell Culture Protocols

The growth conditions of microbial cell cultures and the time of sample collection should be optimized and standardized when growing cells for protein extraction. Because cells may excrete proteases and other extracellular enzymes, and compounds in the medium may interfere with extraction, wash cultures with an isotonic buffer, such as PBS or sucrose before solubilization. - [Read Extraction and Solubilization of Total Protein from Microorganisms Protocol]

Gentle lysis of cells in sucrose-containing Mito-Buffer - http://www.celldeath.de/apometh/mitobuff.html

Category: Cell Biology and Cell Culture: Cell Lysis Protocols

Gentle lysis of mammalian cells for cytochrome c release assay. Based on Yang et al., 1997 Celldeath.de - [Read Gentle lysis of cells in sucrose-containing Mito-Buffer]

Gentle lysis of mammalian cells for cytochrome c release assay - http://www.celldeath.de/apometh/mitobuff.html

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Lysis in the sucrose-containing Buffer ("Mito-Buffer") is supposed to prevent accidential disrupture of the mitochondria to prevent the leakage of mitochondrial proteins (such as cytochrome c) into the cytosol. Celldeath.de - [Read Gentle lysis of mammalian cells for cytochrome c release assay]

Lysosome Isolation in Isotonic Sucrose Protocol - http://homepages.gac.edu/~cellab/chpts/chpt7/ex7-1.html

Category: Cell Organelle Protocols: Lysosome Protocols

Protocol for the isolation of lysosome in isotonic sucrose. - [Read Lysosome Isolation in Isotonic Sucrose Protocol]

Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol - http://www.natureprotocols.com/2006/09/15/northern_blot_analysis_of_mrna.php

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

The protocol consists of a method for the generation of cytoplasmic extracts from mammalian cells (in this case, 293T cells) without the disruption of polyribosomes, the separation of ribosomal components and polyribosomes by sucrose gradient centrifugation, the isolation of mRNA from these fractions, and detection of mRNA by Northern blot analysis. - [Read Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol]

Purification of Bacteriophage lambda Arms: Centrifugation through Sucrose Density Gradients Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3990

Category: Viral Protocols

Protocol is used to prepare the arms of any bacteriophage {lambda} vector. - [Read Purification of Bacteriophage lambda Arms: Centrifugation through Sucrose Density Gradients Protocol]

Sucrose Gradient Protocol - http://www.med.unc.edu/~hdohlman/sucrose.html

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols: Membrane Isolation Protocols

Protocol for sucrose gradient. - [Read Sucrose Gradient Protocol]

Articles (1-1 of 1)

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.