The pH is an important parameter controlling many metabolic and signalling pathways in living cells. Recombinant fluorescent pH indicators (pHluorins) have come into vogue for monitoring cellular pH. They are derived from the most popular Aequorea victoria GFP (Av-GFP). Here, we present a novel fluorescent pH reporter protein from the orange seapen Ptilosarcus gurneyi (Pt-GFP) and compare its properties with pHluorins for expression and use in plants. - [Read A Novel Fluorescent pH Probe for Expression in Plants]
CHO Lec3.2.8.1 cells have 4 independent mutations in the N and O glycosylation pathways. When cultured with alpha-glucosidase I inhibitor N-butyl-deoxynojirimycin, glycoproteins produced in CHO Lec3.2.8.1 cells are completely susceptible to Endo H digestion. Endo H cleaves chitobiose, leaving a single N-linked N-acetylglucosamine per site which is ideal for maintenance of protein solubility and special carb-protein interactions, such as between the first N-acetyl glucosamine residue and tryp. - [Read Establishment of Stable Transfectant of CHO Lec Cells Protocol]
Describes generating CTL against some commonly used target antigens. Two methods for the quantitation of CTL activity are described based on the two pathways used bt CTL to kill target cells. In one pathway, they release lytic granules containing perforin and granzymes, leading to apoptosis and target cell lysis. In a second pathway, they trigger apoptosis via Fas/Fas ligand interactions. - [Read Induction and Measurement of Cytotoxic T Lymphocyte Activity Protocol]
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]
Background and methods to study plant transport. Includes new methods to study protein trafficking in plant cells, includes: Identification of protein sorting pathways in non purified samples; Localization of organelle proteins by isotype tagging/isotype-coded affinity tag; Coupling of chemical genomics and proteomics; Top down mass spectrometry; Compartment-specific markers to aid in the purification of organelles. - [Read Understanding Protein Trafficking in Plant Cells Through Proteomics]