Assays for B Lymphocyte Function Protocols- https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66340AD6485AA0FEB8F538B2FD389E&objectid=6674A538AC9B642C5CD51556EF7B3D26
Describes the antigenic stimulation of in vitro antibody production by B cells and the subsequent measurement of secreted antibodies. The first protocol is a generalized system for inducing in vitro antibody production and can accommodate various types of antigens under study. Secreted antibodies can then be measured with an enzyme-linked immunosorbent assay (ELISA) or other soluble-antibody detection systems. - [Read Assays for B Lymphocyte Function Protocols]
Cytokine sandwich ELISA are sensitive enzyme immunoassays that can specifically detect and quantitate the concentration of soluble cytokine and chemokine proteins. BD Biosciences - [Read Cytokine ELISA Protocol]
Fixation must convert the soluble contents of the cell (ie proteins, carbohydrates, ... http://www.protocol-online.org/prot/Histology/Fixation/ ...General Overview of Histology and Tissue Fixation. BIOS 576. Rice. - [Read General Overview of Histology]
Protocol for the Stimulation of human peripheral blood mononuclear cells with anti-human CD3 monoclonal antibody; MTT assay for detection of cellular proliferation. Human PBMCs can be activated in vitro by soluble anti-human CD3 antibodies. Performed titration studies with these antibodies and established the following protocol for stimulation of PBMC. - [Read Human T Cell Activation Protocol]
Immunoprecipitation / Kinase Assay Protocol. Upstate. Protocol applicable only to kinases whose activity is not altered by cell lysis or immunoprecipitation procedures, and do not require soluble cofactors for activity. - [Read Immunoprecipitation / Kinase Assay]
Protocol describes a nondenaturing immunoprecipitation (IP) for mammalian cells. Prefer to use denaturing IPs to recover labeled proteins from pulse-chase experiments. However, the nondenaturing protocol is useful when one wishes to separate soluble from insoluble proteins, or when the antibody being used recognizes a native epitope. - [Read Nondenaturing Protein Immunoprecipitation from Mammalian Cells Protocol]
Protocol describes how to produce a soluble nuclear extract rich in basal pol II transcription factors from Drosophila embryos. This is a cell-free extract that contains all the necessary transcription factors and is capable of accurate initiation of transcription by RNA polymerase II but is deficient in core histones and histone H1. - [Read Preparation of a Highly Efficient Transcription Extract from Drosophila Embryos Protocol]
Production of T Cell Clones Protocol- https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66347791E1CAD019BE41F3277340CA&objectid=6674BCB8F6BCF7B8032EEBAF7A3D08BD
Provides methods for the derivation of specific types of T cell clones: preparation and maintenance of alloreactive murine helper T (TH) lymphocyte and cytotoxic T lymphocyte (CTL) clones using the limiting dilution technique and derivation of TH clones reactive with soluble protein antigens including a method for the selection of either TH1 or TH2 lymphocyte subsets. - [Read Production of T Cell Clones Protocol]
The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and Cell Cycle Distribution of Primary B Cells Using Propidium Iodide]
The water soluble, DNA intercalator, propidium iodide (PI), is used to bind to DNA after permeabilization of cells with NP40. The amount of dye bound correlates with the content of DNA within a given cell. Once cells are stained, they are analyzed on a flow cytometer. The relative content of DNA indicates the distribution of a population of cells throughout the cell cycle. - [Read Quantification of Apoptosis and the Cell Cycle Distribution of Primary B Cells Using PI]
Protocol for myelin sheath. Luxol Fast Blue is the alcohol soluble counterpart of the water soluble Alcian Blue. Staining is due to lipoproteins, and the mechanism is one of an acid-base reaction with salt formation; the base of the lipoprotein replaces the base of the dye. - [Read Staining for Myelin Sheath Protocol]
In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.