Hu Intracell Staining Starter Kit 25Tst

• 产品编号：BD-559302      品牌：BD-Pharmingen       原厂货号：559302
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

描述：

Improved methods have continually been sought to analyze cytokineproducing
cells at the single cell level. Techniques for analyzing individual
cytokine-producing cells include immunohistochemistry,
immunocytochemistry, ELISPOT, in-situ hybridization, limiting dilution
analysis, and single cell PCR.1 All of these techniques have their respective
advantages, but also significant drawbacks including the requirement for
either technical proficiency or tedious data collection and analysis. Flow
cytometry, however, is a powerful analytical technique in which individual
cells can be simultaneously analyzed for several parameters, including size
and granularity, as well as the expression of surface and intracellular
markers defined by fluorescent antibodies.1-5
Fluorescent anti-cytokine and anti-chemokine monoclonal antibodies have
been used extensively for the intracellular staining and multiparameter flow
cytometric analysis of individual cytokine-producing cells within purified
and mixed cell populations.1,3-8 Multicolor immunofluorescent staining
with antibodies specific for intracellular cytokines and cell surface markers
provides a high resolution method to identify the nature and frequency of
cells that express cytokines in either a restricted (eg, Th1- versus Th2-like
cells) or unrestricted (eg, Th0-like cells) pattern.9,10 While enabling highly
specific and sensitive measurements of several parameters for individual
cells simultaneously, this method also has the capacity for rapid analysis of
large numbers of cells required for making statistically significant
measurements.2
Staining of intracellular cytokines depends on the identification of cytokinespecific
monoclonal antibodies that are compatible with a fixationpermeabilization
procedure.11-13 Optimal intracellular cytokine staining has
been reported using a combination of fixation with paraformaldehyde and
subsequent permeabilization of cell membranes with the detergent saponin.
Paraformaldehyde fixation allows for the preservation of cell morphology
and intracellular antigenicity, while also enabling the cells to withstand
permeabilization by detergent.14 Membrane permeabilization by saponin
allows the fluorochrome-conjugated cytokine-specific monoclonal antibody
to penetrate the cell membrane, cytosol, and membranes of the endoplasmic
reticulum and Golgi apparatus. Critical parameters for intracellular cytokine
staining include the following: cell type; activation protocol and cellular
response kinetics (important for determining when to harvest cells); the
inclusion of a protein transport inhibitor during cell activation; and the
choice of an anti-cytokine antibody. Lack of information concerning these
parameters has often precluded researchers from utilizing the intracellular
staining technique.

原厂资料：

注意事项：

This kit contains the essential reagents for the activation of a human
peripheral blood cell population and their subsequent intracellular staining
to detect cytokine-producing cells. Positive and negative controls are
included to assist the investigator in utilizing this procedure. The reagents
provided in the Starter Kit are described below.