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AB236210

Respiratory Burst Assay Kit (Neutrophil/Monocyte)

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(4 Publications)

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.
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Functional Studies - Respiratory Burst Assay Kit (Neutrophil/Monocyte) (AB236210)
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Supplier Data

Functional Studies - Respiratory Burst Assay Kit (Neutrophil/Monocyte) (AB236210)

Flow cytometric analysis of human peripheral blood.

Human peripheral blood was treated with Dihydrorhodamine 123 Assay Reagent followed by stimulation with PMA for 45 minutes to induce the respiratory burst. The RBC were lysed and the leukocytesanalysed by flow cytometry. Left panel : Forwardangle light scatter(FSC) and side scatter (SSC) segregate neutrpgils (gate P1), monocytes (gate P2) and lymphocytes (gate P3) for susequent analysis in the other panels.
Remaining panels : Enhanced oxidation of Dihydrorhodamine 123 Assay Reagent to rhodamine 123 is indicated by a right shift in the x-axis FL1 fluorescence in the gated neutrophils and monocytes, but not in the lymphocytes. The black lines represent untreated cells and the red lines represent PMA-treated cells.

Functional Studies - Respiratory Burst Assay Kit (Neutrophil/Monocyte) (AB236210)
  • FuncS

Supplier Data

Functional Studies - Respiratory Burst Assay Kit (Neutrophil/Monocyte) (AB236210)

Flow cytometric analysis of mouse neutrophil respiratory burst.

Mouse bone marrow cells (top) and caesi-induced peritoneal exudate cells (bottom) were treated with Dihydrorhodamine 123 Assay Reagent followed by stimulation with PMA (100 nM) for 45 minutes to induce the respiratory burst. The RBC were lysed and the leukocytes analyzed by flow cytometry. Left panels : The neutrophils are identified by an intermediate forward angle light scatter (FSC) and high side scatter (SSC), and are gated (P1) for subsequent analysis in the other panels. Right panels : Enhanced oxidation of Dihydrorhodamine 123 Assay Reagent to rhodamine 123 is indicated by a right shift in the x-axis FL1 fluorescence in the gated neutrophils from samples treated with PMA (red line) versus untreated cells (black line).

Functional Studies - Respiratory Burst Assay Kit (Neutrophil/Monocyte) (AB236210)
  • FuncS

Supplier Data

Functional Studies - Respiratory Burst Assay Kit (Neutrophil/Monocyte) (AB236210)

PMA dose-response.

Human peripheral blood was treated with Dihydrorhodamine 123 Assay Reagent followed by stimulation with the indicated concentrations of PMA for 45 minutes to induce the respiratory burst. The RBC were lysed and the leukocytes analyzed by flow cytometry.

Key facts

Detection method

Flow cytometry-fluorescent

Sample types

Suspension cells, Adherent cells, Whole Blood

Assay Platform

Flow cytometer

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Reagents, Controls & Accessories

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Red Blood Cell (RBC) Lysis Buffer (10x)

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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.
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What's included?

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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
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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.
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Product protocols

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Target data

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Publications (4)

Recent publications for all applications. Explore the full list and refine your search

Nature communications 16:4945 PubMed40436823

2025

WNK1 mediates M-CSF-induced macropinocytosis to enforce macrophage lineage fidelity.

Applications

Unspecified application

Species

Unspecified reactive species

Alissa J Trzeciak,Zong-Lin Liu,Mohamed Gatie,Adam S Krebs,Waleska Saitz Rojas,Anya J O'Neal,Ann K Baako,Zhaoquan Wang,Justin Nelson,Isabella C Miranda,Jazib Uddin,Allie Lipshutz,Jian Xie,Chou-Long Huang,Pedro H V Saavedra,Anna-Katerina Hadjantonakis,Michael Overholtzer,Michael S Glickman,Arohan R Subramanya,Thomas Vierbuchen,Jon Iker Etchegaray,Christopher D Lucas,Christopher N Parkhurst,Justin S A Perry

Frontiers in cellular and infection microbiology 14:1394070 PubMed38895731

2024

Probiotic bacteria can modulate immune responses to paratuberculosis vaccination.

Applications

Unspecified application

Species

Unspecified reactive species

Maddi Oyanguren,Elena Molina,Maitane Mugica,Iraia Ladero-Auñon,Miguel Fuertes,Miguel Fernández,Julio Benavides,Natalia Elguezabal

Nanomedicine : nanotechnology, biology, and medicine 54:102712 PubMed37838100

2023

Liposomes - Human phagocytes interplay in whole blood: effect of liposome design.

Applications

Unspecified application

Species

Unspecified reactive species

Miriam Giambelluca,Elena Markova,Claire Louet,Bjørg Steinkjer,Rune Sundset,Nataša Škalko-Basnet,Sjoerd Hak

Frontiers in immunology 11:586685 PubMed33042165

2020

Gut Ischemia Reperfusion Injury Induces Lung Inflammation via Mesenteric Lymph-Mediated Neutrophil Activation.

Applications

Unspecified application

Species

Unspecified reactive species

Yonggang Ma,Taylor Zabell,Alexandra Creasy,Xiaoyuan Yang,Victor Chatterjee,Nuria Villalba,Erik B Kistler,Mack H Wu,Sarah Y Yuan
View all publications
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