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Human NOX2/gp91phox ELISA Kit- Intracellular is a single-wash 90-min Simplestep used to quantify Human NOX2/gp91phox with a sensitivity of 1.24 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

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Images

Sandwich ELISA - Human NOX2/gp91phox ELISA Kit- Intracellular (AB300334), expandable thumbnail
  • Sandwich ELISA - Human NOX2/gp91phox ELISA Kit- Intracellular (AB300334), expandable thumbnail

Key facts

Detection method
Colorimetric
Sample types
Cell Lysate
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
6.25 - 400 pg/mL
Assay time
1h 30m
Sensitivity
= 1.24 pg/mL

Reactivity data

Application
sELISA
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

Critical component of the membrane-bound oxidase of phagocytes that generates superoxide. It is the terminal component of a respiratory chain that transfers single electrons from cytoplasmic NADPH across the plasma membrane to molecular oxygen on the exterior. Also functions as a voltage-gated proton channel that mediates the H(+) currents of resting phagocytes. It participates in the regulation of cellular pH and is blocked by zinc.

Alternative names

What's included?

96 Tests
Components
10X Human NOX2/gp91phox Capture Antibody
1 x 600 µL
10X Human NOX2/gp91phox Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
5X Cell Extraction Buffer PTR (ab193970)
1 x 10 mL
Antibody Diluent CPR2
1 x 6 mL
Human NOX2/gp91phox Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
1 x 12 mL
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 96 Wells
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

Recommended products

Human NOX2/gp91phox ELISA Kit- Intracellular is a single-wash 90-min Simplestep used to quantify Human NOX2/gp91phox with a sensitivity of 1.24 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

Key facts

Detection method
Colorimetric
Sample types
Cell Lysate
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
6.25 - 400 pg/mL
Assay time
1h 30m
Assay Platform
Pre-coated microplate (12 x 8 well strips)
Sensitivity
= 1.24 pg/mL
General recovery
101%

Precision

Intra assay

Sample
Extract
n
8
mean
-
SD
-
C.V.
9.4

Inter assay

Sample
Extract
n
3
mean
-
SD
-
C.V.
10.1

Recovery

Sample specific recovery

Sample type
Cell Lysate
Average %
= 101
Range
98 - 107 %

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Notes

Human NOX2/gp91phox SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of NOX2/gp91phox protein in human cell extract. Quantitate Human NOX2/gp91phox with 1.24 pg/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

NOX2 also known as gp91phox or cybb is a membrane-bound protein that plays an important role in the production of reactive oxygen species (ROS). This protein functions as a catalytic component of the NADPH oxidase complex which is primarily found in phagocytes like neutrophils and macrophages. The NOX2 protein has a molecular weight of approximately 91 kDa. It is expressed prominently in the cell membranes of these immune cells where it participates in the body's defense mechanisms.

Biological function summary

NOX2 participates in the generation of superoxide by using NADPH as a substrate an essential function performed by the multi-subunit NADPH oxidase complex. This process helps in the formation of microbicidal ROS which neutrophils and macrophages require for killing pathogens. The gp91phox component NOX2's alternate name partners with p22phox and other cytosolic subunits like p47phox and p67phox to form the fully active enzyme complex. This emphasizes NOX2's role in the immune system's oxidative burst an important antimicrobial response.

Pathways

NOX2 is an integral part of the oxidative burst pathway important in host defense. Its activity connects with the MAPK signaling pathway which can be activated by the presence of ROS produced by NOX2. This pathway involves proteins such as ERK JNK and p38 MAPKs which are related through the signaling events that lead to diverse cellular responses including inflammation and stress responses.

Associated diseases and disorders

NOX2 dysregulation is associated with chronic granulomatous disease (CGD) a condition characterized by a compromised ability of phagocytes to produce ROS leading to recurrent infections. Additionally NOX2 activity links with cardiovascular diseases where its overproduction of ROS can cause oxidative stress contributing to atherosclerosis. The functioning of p22phox another protein in the NADPH oxidase complex becomes particularly relevant in these contexts as it partners with NOX2 to generate the harmful superoxide in such diseases.

Product promise

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2 product images

  • Sandwich ELISA - Human NOX2/gp91phox ELISA Kit- Intracellular (ab300334), expandable thumbnail

    Sandwich ELISA - Human NOX2/gp91phox ELISA Kit- Intracellular (ab300334)

    Example of human NOX2/gp91phox standard curve. Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - Human NOX2/gp91phox ELISA Kit- Intracellular (ab300334), expandable thumbnail

    Sandwich ELISA - Human NOX2/gp91phox ELISA Kit- Intracellular (ab300334)

    Interpolated concentration of native NOX2/gp91phox was measured in duplicate at different sample concentrations. Undiluted samples are as follows: PBMC cell extract 145 µg/mL and HL-60 cell extract 250 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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