Antigen Design and Sera Purification. Custom antisera. Sigma Aldrich. Peptide Selection and Design, Coupling Strategy
# Selecting the Protein Carrier, Multiple Antigenic Peptides (MAPs), Choice of Host, Adjuvant, Immunization, & Sera Collection, Antisera Purification, Ammonium Sulfate Precipitation, Protein A/G, Immunoaffinity Purification. - [Read Antigen Design and Sera Purification]
Biological Buffers a reference- http://www.sigmaaldrich.com/Brands/Fluka___Riedel_Home/Bioscience/BioChemika_Ultra/Biological_Buffers.html
ECACC Handbook of Cell Culture. Fundamental Techniques in Cell Culture: A Labratory Handbook. Sigma Aldrich. Excellent Guide to Cell Culture, aseptic technique, storage of cell lines, subculture of cells. - [Read Handbook of Cell Culture. - Sigma Aldrich.]
Several common drugs, their targets, and protocols are described for studying organelle distribution and trafficking. The drugs are readily available from general suppliers, including Sigma, Roche, and Calbiochem. - [Read Membrane Trafficking and Organelle Reagents]
Peptide Handling and Storage- http://www.sigmaaldrich.com/Brands/Sigma_Genosys/Custom_Peptides/Key_Resources/Handling___Storage.html
In general, peptide solutions are stable for up to a week at 4°C. However, if the peptide sequence has inherent instability (Peptide Stability), it might be better to freeze the solution when not in use. Peptide solutions at pH>8 should also be frozen when not in use. Sigma Aldrich. - [Read Peptide Handling and Storage]
Peptide Handling Guides. Storage Guidelines for Lyophilized Peptides, Strategy for Dissolving Single Peptides, Determining Solubility Characteristics of peptides, Dissolving Approach for Charged Peptides, Dissolving Approach for Hydrophobic/Uncharged Peptides, Guidelines for Dissolving Several Peptides, Peptide Stability and Potential Degradation Pathways, and storage of peptides. Sigma Aldrich. PDF. - [Read Peptide Handling Guides]
The Cell Growth Environment- http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook/Cell_Culture_Techniques_6.html#Environment
Cultured cells require a sterile environment and a supply of nutrients for growth. Basic Constituents of media. Sigma - [Read The Cell Growth Environment]
Treatment of Cells with 5-aza-dC. protocol PDF- http://www.shmu.edu.cn/courses/2005aut/upload/20051116/Hongmei%20Xu%202003%20CANCER%20RESEARCH%20%20Aberrant%20Methylation%20and%20Silencing%20of%20ARHI,%20an%20Imprinted%20Tumor%20Suppressor%20Gene%20in%20which%20the%20Function%20Is%20Lost%20in%20Breast%
"Cells were seeded at a density of 1X106 cells/100-mm dish with 10% FBS and
allowed to attach over a 24-h period. 5-Aza-dC (Sigma) was then added to a
final concentration of 0.2–1 M, and the cells were allowed to grow for 5 days. The medium with or w - [Read Treatment of Cells with 5-aza-dC. protocol PDF]
DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.
This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.
Histone H1 Kinase Activity Assay Protocol. This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography.