What is intracellular cytokine staining?
What is intracellular cytokine staining?
Intracellular cytokine staining (ICS) is a method of cytokine analysis that provides information on the type of cytokines that are produced upon (antigen-specific) stimulation of T cells and B cells. Cytokine production can then be assessed via flow cytometry.
What is the function of T-bet?
T-bet is an essential regulator of effector differentiation and function in multiple different immune lineages, including CD4, CD8, NKT, ILC (including NK) and B cells. Inflammatory cytokines induce T-bet expression in a graded manner, which in turn regulates distinct differentiation outcomes.
What cells express t-bet?
T-bet is also expressed in a subset of T-cell lymphomas, particularly those that express other markers of Th1 T cell differentiation, and in a subset of B-cell non-Hodgkin’s lymphomas.
What is blimp1?
BLIMP-1 helps the production of short-lived effector T cells and clonally exhausted T cells. It also helps with the migration of T cells out of the spleen and lymph nodes into peripheral tissues. However, BLIMP-1 does not promote the production of long-lived effector memory cells.
Do NK cells express T-bet?
The bulk of mature murine (6, 8, 9) and human (10–12) NK cells express high levels of T-bet and Eomes, but until recently, their impact on NK cell function was not known.
What does T-bet mean?
T-box expressed
T-box transcription factor TBX21, also called T-bet (T-box expressed in T cells) is a protein that in humans is encoded by the TBX21 gene.
Is T-bet a transcription factor?
T-bet (Tbx21) is an immune cell transcription factor originally described as the master regulator of Th1 cell development, although is now recognized to have a role in both the adaptive and innate immune systems. T-bet also directs T-cell homing to pro-inflammatory sites by the regulation of CXCR3 expression.
Is ELISpot more sensitive than ELISA?
An ELISPOT assay can 100 to 400 times more sensitive than a conventional ELISA because the secreted protein is captured directly onto the well of an ELISPOT plate before having the chance to be diluted in the culture supernatant, degraded by proteases or captured by receptors on adjacent cells.
What causes background in ELISpot?
High background. Wash both sides of the membrane with distilled water before and after color development. Some reagents may leak through the membrane into the base of the plate, and these can cause high background if not washed away.
Intracellular cytokine staining (ICS) is a very useful and widely used flow cytometry based assay which detects the production and accumulation of cytokines within the endoplasmic reticulum after cell stimulation.
What is the intacellular flow cytometry staining protocol?
The Intacellular Flow Cytometry Staining Protocol describes the process for intracellular staining of various cell types (in vivo-stimulated tissues, in vitro-stimulated cultures, and whole blood) for flow cytometry using BioLegend’s proprietary buffers and antibodies.
Can intracellular cytokines and cell surface markers be measured simultaneously?
Microscopic and flow cytometric analysis showed a highly significant correlation. Using three-color flow cytometry it was possible to measure intracellular cytokines and cell surface markers simultaneously.
How do you stain intracellular antigens in centrifuge?
Centrifuge at 350xg for 5 minutes and discard the supernatant. Wash cells 1X with Cell Staining Buffer and perform cell surface immunofluorescent staining as described above. Fix, permeabilize, and stain intracellular antigens as described above. Set PMT voltage and compensation using cell surface staining controls.
