JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB274401

Human NLRP3 ELISA Kit

3

(1 Review)

|

(10 Publications)

Human NLRP3 ELISA Kit is a single-wash 90-min Simplestep used to quantify Human NLRP3 with a sensitivity of 1.63 ng/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
- Cited in over 10 citations

View Alternative Names

C1orf7, CIAS1, NALP3, PYPAF1, NLRP3, Angiotensin/vasopressin receptor AII/AVP-like, Caterpiller protein 1.1, Cold-induced autoinflammatory syndrome 1 protein, Cryopyrin, PYRIN-containing APAF1-like protein 1, CLR1.1

3 Images
ELISA - Human NLRP3 ELISA Kit (AB274401)
  • ELISA

Supplier Data

ELISA - Human NLRP3 ELISA Kit (AB274401)
Sandwich ELISA - Human NLRP3 ELISA Kit (AB274401)
  • sELISA

Supplier Data

Sandwich ELISA - Human NLRP3 ELISA Kit (AB274401)

Example of human NLRP3 standard curve in 1X Cell Extraction Buffer PTR.

NLRP3 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human NLRP3 ELISA Kit (AB274401)
  • sELISA

Supplier Data

Sandwich ELISA - Human NLRP3 ELISA Kit (AB274401)

Interpolated concentrations of native NLRP3 in human THP-1 and liver samples based on a 62.5 μg/mL and 500 µg/mL extract load, respectively.

The concentrations of NLRP3 were measured in duplicate and interpolated from the NLRP3 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean NLRP3 concentration was determined to be 8.7 ng/mL in THP-1 cell extract and undetectable in Liver Extract.

Key facts

Detection method

Colorimetric

Sample types

Tissue Extracts, Cell Lysate

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 1.63 ng/mL

Range

4.38 - 280 ng/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
style
left-column

Product details

Human NLRP3 ELISA Kit ab274401 is a rapid single-wash 90-min Sandwich ELISA to measure Human NLRP3 in cell lysate, tissue extracts. This SimpleStep sensitivity is 1.63 ng/mL.

How the assay works

Human NLRP3 SimpleStep ELISA®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.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human NLRP3 ELISA Kit ab274401 protocol summary

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-NLRP3 antibody [EPR23094-118] - BSA and Azide free (Capture) ab274451
- Anti-NLRP3 antibody [EPR23094-101] - BSA and Azide free (Detector) ab274451

NACHT, LRR and PYD domains-containing protein 3 (NLRP3), encoded by the NLRP3 gene, is known to have a vital role in the innate immunity and inflammation. NLRP3 is highly expressed in macrophages and as a component of the inflammasomes in which it functions to detect products of damaged cells and subsequently activates itself to trigger an immune response. Mutations in the NLRP3 has been shown in studies to be associated with a wide spectrum of dominantly inherited autoimmune diseases. The standard protein provided is an N-terminal fragment of human NLRP3 that has been mass calibrated to full-length recombinant NLRP3.

REACH authorisation
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.

style
left-column

Precision

[ { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "5.1" }, { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.5" } ]
style
left-column

Recovery

[ { "sample": "Tissue Extracts", "range": "110 - 113 %", "average": "= 112" }, { "sample": "Cell Lysate", "range": "111 - 112 %", "average": "= 112" } ]
style
left-column

What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab274401-1x96Tests", "assetComponentDetails": [ { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB274401-CMP06", "productcode":"" }, { "size":"1 x 12 mL", "name":"Sample Diluent NS (ab193972)", "number":"AB274401-CMP12", "productcode":"" }, { "size":"1 x 6 mL", "name":"Antibody Diluent CPR2", "number":"AB274401-CMP13", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB274401-CMP11", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human NLRP3 Lyophilized Recombinant Protein", "number":"AB274401-CMP08", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human NLRP3 Capture Antibody", "number":"AB274401-CMP03", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Development Solution", "number":"AB274401-CMP07", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human NLRP3 Detector Antibody", "number":"AB274401-CMP04", "productcode":"" }, { "size":"1 x 10 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB274401-CMP09", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seals", "number":"AB274401-CMP10", "productcode":"" }, { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB274401-CMP05", "productcode":"" } ] } } }
style
left-column

Properties and storage information

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

Supplementary information

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

NLRP3 also known as NALP3 or cryopyrin is a protein important for the inflammatory response. It is a component of the inflammasome which senses pathogenic microorganisms and stress. NLRP3 weighs around 118 kDa and is widely expressed in innate immune cells particularly macrophages and dendritic cells. Researchers often use techniques such as 'NLRP3 WB' (Western blot) and 'NLRP3 immunofluorescence' to study NLRP3 expression and localization.
Biological function summary

NLRP3 is central to the body's defense mechanism by participating in forming the inflammasome complex which activates caspase-1. This activation leads to the cleavage of pro-inflammatory cytokines like IL-1β and IL-18 into their active forms. NLRP3's function is essential for mounting an effective immune response to infections and damage signals. It senses a range of stimuli including crystalline substances and cellular stress triggering an immune response. Its expression is tightly regulated to prevent excessive inflammation.

Pathways

The NLRP3 inflammasome is key in the innate immune response and inflammation pathways. It links cellular stress signals to inflammation through the activation of caspase-1 which is important for the IL-1 signaling pathway. NLRP3 interacts with other proteins like ASC (apoptosis-associated speck-like protein containing a CARD) to form the inflammasome complex which is a core part of the inflammatory signaling pathways and links innate and adaptive immunity.

NLRP3 has a significant role in inflammatory diseases and is linked to conditions like cryopyrin-associated periodic syndromes (CAPS) and gout. Mutations or dysregulation of NLRP3 lead to hyperactivation resulting in autoimmune and autoinflammatory conditions. NLRP3's interaction with IL-1 contributes to the excessive inflammatory response seen in these disorders. Understanding NLRP3's function in these pathways can inform therapeutic approaches targeting chronic inflammatory diseases.
style
left-column

Product protocols

style
left-column

Target data

Sensor component of the NLRP3 inflammasome, which mediates inflammasome activation in response to defects in membrane integrity, leading to secretion of inflammatory cytokines IL1B and IL18 and pyroptosis (PubMed : 16407889, PubMed : 18403674, PubMed : 18604214, PubMed : 23582325, PubMed : 25686105, PubMed : 27929086, PubMed : 28656979, PubMed : 28847925, PubMed : 30487600, PubMed : 30612879, PubMed : 31086327, PubMed : 31086329, PubMed : 31189953, PubMed : 33231615, PubMed : 34133077, PubMed : 34341353, PubMed : 34512673, PubMed : 36442502). In response to pathogens and other damage-associated signals that affect the integrity of membranes, initiates the formation of the inflammasome polymeric complex composed of NLRP3, CASP1 and PYCARD/ASC (PubMed : 16407889, PubMed : 18403674, PubMed : 27432880, PubMed : 28847925, PubMed : 31189953, PubMed : 33231615, PubMed : 34133077, PubMed : 34341353, PubMed : 36142182, PubMed : 36442502). Recruitment of pro-caspase-1 (proCASP1) to the NLRP3 inflammasome promotes caspase-1 (CASP1) activation, which subsequently cleaves and activates inflammatory cytokines IL1B and IL18 and gasdermin-D (GSDMD), promoting cytokine secretion and pyroptosis (PubMed : 23582325, PubMed : 28847925, PubMed : 31189953, PubMed : 33231615, PubMed : 34133077, PubMed : 34341353). Activation of NLRP3 inflammasome is also required for HMGB1 secretion; stimulating inflammatory responses (PubMed : 22801494). Under resting conditions, ADP-bound NLRP3 is autoinhibited (PubMed : 35114687). NLRP3 activation stimuli include extracellular ATP, nigericin, reactive oxygen species, crystals of monosodium urate or cholesterol, amyloid-beta fibers, environmental or industrial particles and nanoparticles, such as asbestos, silica, aluminum salts, cytosolic dsRNA, etc (PubMed : 16407889, PubMed : 18403674, PubMed : 18604214, PubMed : 19414800, PubMed : 23871209). Almost all stimuli trigger intracellular K(+) efflux (By similarity). These stimuli lead to membrane perturbation and activation of NLRP3 (By similarity). Upon activation, NLRP3 is transported to microtubule organizing center (MTOC), where it is unlocked by NEK7, leading to its relocalization to dispersed trans-Golgi network (dTGN) vesicle membranes and formation of an active inflammasome complex (PubMed : 36442502). Associates with dTGN vesicle membranes by binding to phosphatidylinositol 4-phosphate (PtdIns4P) (PubMed : 30487600, PubMed : 34554188). Shows ATPase activity (PubMed : 17483456).. Independently of inflammasome activation, regulates the differentiation of T helper 2 (Th2) cells and has a role in Th2 cell-dependent asthma and tumor growth (By similarity). During Th2 differentiation, required for optimal IRF4 binding to IL4 promoter and for IRF4-dependent IL4 transcription (By similarity). Binds to the consensus DNA sequence 5'-GRRGGNRGAG-3' (By similarity). May also participate in the transcription of IL5, IL13, GATA3, CCR3, CCR4 and MAF (By similarity).
See full target information NLRP3
style
left-column

Publications (10)

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

Cardiovascular diabetology 24:203 PubMed40361188

2025

Diabetes mellitus aggravates myocardial inflammation and oxidative stress in aortic stenosis: a mechanistic link to HFpEF features.

Applications

Unspecified application

Species

Unspecified reactive species

Melissa Herwig,Marcel Sieme,Andrea Kovács,Muchtiar Khan,Andreas Mügge,Wolfgang E Schmidt,Ferhat Elci,Shan Sasidharan,Peter Haldenwang,Jan Wintrich,Benjamin Sasko,Ibrahim Akin,Máthé Domokos,Francesco Paneni,Ibrahim El-Battrawy,Zoltán V Varga,Francisca Saraiva,Adelino F Leite-Moreira,Péter Ferdinandy,Loek van Heerebeek,Inês Falcão-Pires,Nazha Hamdani

Advances in clinical and experimental medicine : official organ Wroclaw Medical University 34:1459-1466 PubMed39629849

2024

NLRP3 inflammasome in expressed prostatic secretions as a potential biomarker of chronic prostatitis/chronic pelvic pain syndrome.

Applications

Unspecified application

Species

Unspecified reactive species

Chao-Guang Ma,Ying-Nan Liu,Hua-Dong Wang

Molecular medicine reports 30: PubMed38873985

2024

Hydrogen sulfide mitigates ox‑LDL‑induced NLRP3/caspase‑1/GSDMD dependent macrophage pyroptosis by S‑sulfhydrating caspase‑1.

Applications

Unspecified application

Species

Unspecified reactive species

Zhenli Jia,Xulin Zhang,Zhiyi Li,Hanyu Yan,Xiangqin Tian,Chenghua Luo,Ketao Ma,Ling Li,Liang Zhang

Journal of molecular endocrinology 73: PubMed38722222

2024

LIF-STAT signaling in decidual cells: a possible role in embryo implantation and early pregnancy.

Applications

Unspecified application

Species

Unspecified reactive species

Hsien-Ming Wu,Liang-Hsuan Chen,Wei-Jung Chiu,Chia-Lung Tsai

Journal of periodontal research 59:204-219 PubMed37957813

2023

Gallic acid induces osteoblast differentiation and alleviates inflammatory response through GPR35/GSK3β/β-catenin signaling pathway in human periodontal ligament cells.

Applications

Unspecified application

Species

Unspecified reactive species

Thi Hoa Pham,Eun-Nam Kim,Nguyen Minh Trang,Gil-Saeng Jeong

Theranostics 13:531-542 PubMed36632236

2023

SIRT3 mediates the effects of PCSK9 inhibitors on inflammation, autophagy, and oxidative stress in endothelial cells.

Applications

Unspecified application

Species

Unspecified reactive species

Nunzia D'Onofrio,Francesco Prattichizzo,Raffaele Marfella,Celestino Sardu,Elisa Martino,Lucia Scisciola,Lorenza Marfella,Rosalba La Grotta,Chiara Frigé,Giuseppe Paolisso,Antonio Ceriello,Maria Luisa Balestrieri

Cells 12: PubMed36611853

2022

Enhanced Activity of NLRP3 Inflammasome in the Lung of Patients with Anti-Synthetase Syndrome.

Applications

Unspecified application

Species

Unspecified reactive species

Espiridión Ramos-Martinez,Angel E Vega-Sánchez,Gloria Pérez-Rubio,Mayra Mejia,Ivette Buendía-Roldán,Montserrat I González-Pérez,Heidegger N Mateos-Toledo,Warrison A Andrade,Ramcés Falfán-Valencia,Jorge Rojas-Serrano

Frontiers in immunology 13:1064303 PubMed36505450

2022

Persistent expression of NLRP3 in spinal microglia promotes development of lumbar disc degeneration.

Applications

Unspecified application

Species

Unspecified reactive species

Peng Wang,Jing Zhang

Pharmaceutics 14: PubMed35745744

2022

Extracellular Vesicle-Associated MicroRNA-138-5p Regulates Embryo Implantation and Early Pregnancy by Adjusting GPR124.

Applications

Unspecified application

Species

Unspecified reactive species

Hsien-Ming Wu,Tzu-Chi Lo,Chia-Lung Tsai,Liang-Hsuan Chen,Hong-Yuan Huang,Hsin-Shih Wang,John Yu

Inflammation 44:1865-1877 PubMed33860869

2021

Induction of Exaggerated Cytokine Production in Human Peripheral Blood Mononuclear Cells by a Recombinant SARS-CoV-2 Spike Glycoprotein S1 and Its Inhibition by Dexamethasone.

Applications

Unspecified application

Species

Unspecified reactive species

Olumayokun A Olajide,Victoria U Iwuanyanwu,Izabela Lepiarz-Raba,Alaa A Al-Hindawi
View all publications
style
left-column
style
left-column
websiteProtocolBooklet
en
style
right-column

Complementary Products

Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Assay Kits

AB215539

Human IL-18 ELISA Kit

Recombinant|SimpleStep

ELISA - Human IL-18 ELISA Kit (AB215539)

  • 5
  • 1
Assay Kits

AB272463

Human GSDMD ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human GSDMD ELISA Kit (AB272463)

  • 0
  • 0
Primary Antibodies

AB9485

Anti-GAPDH antibody - Loading Control

BOND RX™ Validated|Lab Essentials

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (AB9485)

  • 5
  • 88

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com