Description
Key facts
Detection method:Flow cytometry-fluorescent,
Sample types:Suspension cells, Adherent cells, Whole Blood,
Assay Platform:Flow cytometer
Product details:
Respiratory Burst Assay Kit (Neutrophil/Monocyte) (ab236210) provides PMA, dihydrorhodamine 123, and additional reagents necessary for inducing and quantifying a respiratory burst response in neutrophils and monocytes by flow cytometry. The assay can be performed on whole blood or on cells in various types of cell culture media. The dihydrorhodamine 123 used in this assay, a cell-permeable, non-fluorescent dye, is converted to the fluorescent compound rhidamine 123 by reactive species produced by activated phagocytes to destroy invading microorganisms. The assay has been validated in human and mouse whole blood, or using other sources of leukocytes. Because the assay reagents are not species-specific, this assay can be used in any species or cell type capable of producing a NADPH oxidase-dependent respiratory burst response.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
The respiratory burst also known as oxidative burst is a rapid release of reactive oxygen species (ROS) such as superoxide and hydrogen peroxide. This process occurs primarily in phagocytic cells like neutrophils and monocytes following their activation by pathogens. The mass of the specific enzymes involved such as NADPH oxidase is not a singular number since it consists of a complex (several subunits each with varying mass). Expression of these enzymes is mainly found in neutrophils monocytes and other phagocytic cells where they play a critical role in the host defense mechanism.
Biological function summary
The respiratory burst facilitates the destruction of microbial invaders inside phagocytic cells. This process is independent yet complementary to the lysosomal degradation in these cells. ROS generated during the oxidative burst participate in direct microbial killing and enhancing signaling pathways that further stimulate immune responses. The NADPH oxidase complex catalyzes the reduction of oxygen to superoxide which functions as a precursor for other ROS. This enzymatic machinery is important for efficient pathogen eradication and is integral to the innate immune response.
Pathways
The respiratory burst intertwines with the pathways related to immune responses and inflammation. One critical pathway is the MAPK signaling pathway which activates the burst in response to pathogen recognition. Proteins such as protein kinase C (PKC) assist in regulating the activation of NADPH oxidase during this process. Additionally the respiratory burst contributes to signaling cascades that lead to cytokine production further linking it to the inflammatory response network.
Disruptions in the respiratory burst can lead to conditions like chronic granulomatous disease (CGD) and rheumatoid arthritis. CGD involves mutations in any of the genes responsible for NADPH oxidase functioning resulting in compromised microbial killing. This disorder highlights the importance of an effective oxidative burst in host defense. On the other hand excessive ROS production has been implicated in inflammatory disorders like rheumatoid arthritis where it aggravates tissue damage. In these contexts proteins like TNF-alpha involved in inflammation are often linked to the dysregulated activity of the oxidative burst.
