Name: Anti-NLRP3 antibody [EPR23094-1]
SKU: ab263899
Rating: 4 (2 reviews)

Anti-NLRP3 antibody [EPR23094-1] ab263899 is a rabbit monoclonal antibody that is used in NLRP3 western blotting and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with NLRP3 knockout cell line validation
- Antibody clone EPR23094-1 is cited in over 460 publications

Images

Western blot - Anti-NLRP3 antibody [EPR23094-1] (AB263899), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	I-ELISA	WB	IHC-P	ICC/IF	Flow Cyt (Intra)
Human	Tested	Expected	Tested	Not recommended	Not recommended	Tested
Mouse	Tested	Expected	Tested	Not recommended	Not recommended	Expected
Rat	Expected	Expected	Tested	Not recommended	Not recommended	Expected
Recombinant full length protein	Not recommended	Tested	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes -
Species Human	Dilution info 1/30	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Recombinant full length protein	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Human, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Stimulation may be required to allow detection of the NLRP3 protein in most samples. There may be multiple bands of NLRP3 in WB assay due to different isoforms and PTMs.
Species Rat	Dilution info 1/1000	Notes Stimulation may be required to allow detection of the NLRP3 protein in most samples. There may be multiple bands of NLRP3 in WB assay due to different isoforms and PTMs.
Species Human	Dilution info 1/1000	Notes Stimulation may be required to allow detection of the NLRP3 protein in most samples. There may be multiple bands of NLRP3 in WB assay due to different isoforms and PTMs.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human, Recombinant full length protein	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Rat, Recombinant full length protein	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein	Dilution info -	Notes -

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

See full target information NLRP3

Function

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).

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR23094-1
Purification technique: Affinity purification Protein A
Specificity: NLRP3 is related with inflammation and is cell-type dependent. This antibody detects no signal or very weak signal in most non-diseased tissue samples and untreated cell lysates. Stimulation may be required to allow detection of the NLRP3 protein in most samples, as described in the literature (PMID: 29070054, 29434227, and 22569257).

FURTHER INFORMATION ON SPECIFICITY (Chinese Version) available under the support & downloads section.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-NLRP3 antibody [EPR23094-1] (ab263899) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), I-ELISA, IP and WB.

Anti-NLRP3 antibody [EPR23094-1] (ab263899) was first used in a scientific publication in 2020 and has been cited over 253 times in peer reviewed journals. It's performance in Western blot in human, mouse and rat samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-NLRP3 antibody [EPR23094-1] (ab263899) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-NLRP3 antibody [EPR23094-1] (ab263899) has been confirmed by Western Blot testing in NLRP3 knockout THP-1 cells (Human NLRP3 knockout THP-1 cell line ab280063).

Anti-NLRP3 antibody [EPR23094-1] (ab263899) specifically detects NLRP3 (UniProt ID: Q96P20; Molecular weight: 119kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPR23094-1 - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free ab264468.

NLRP3 inflammasome plays a crucial role in immunology and disease by regulating innate immune responses. It contributes to the development of various immune and inflammation-related diseases, including arthritis, Alzheimer's disease and inflammatory bowel disease. EPR23094-1 is one of the top cited clones on the market with over 527 citations

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary: 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.
Associated diseases and disorders: 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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

12 product images

Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
False colour image of Western blot: Anti-NLRP3 antibody [EPR23094-1] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab263899 was shown to bind specifically to NLRP3. A band was observed at 118 kDa in wild-type THP-1 cell lysates with no signal observed at this size in NLRP3 knockout cell line Human NLRP3 knockout THP-1 cell line ab280063 (knockout cell lysate Human NLRP3 knockout THP-1 cell lysate ab280122). To generate this image, wild-type and NLRP3 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution.
All lanes: Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/500 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: NLRP3 knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human NLRP3 knockout THP-1 cell line (Human NLRP3 knockout THP-1 cell line ab280063)
Performed under reducing conditions.
Predicted band size: 118 kDa
Observed band size: 118 kDa
This image is courtesy of Benedikt Saller, from Prof. Dr. Olaf Groß’ lab at the University of Freiburg.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
Blocking step: 2% SMP in PBS/T for 60 minutes at RT.
All lanes: Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution
Lane 1: Wild-type mouse BMDCs treated with 50 ng/ml LPS at 25000 Cells
Lane 2: NLRP3 knockout mouse BMDCs treated with 50 ng/ml LPS at 25000 Cells
Lane 3: Wild-type human THP1 cells treated with 200 ng/ml LPS at 25000 Cells
Secondary
All lanes: Goat anti-Rabbit HRP at 1/5000 dilution
Developed using the ECL technique.
Predicted band size: 118 kDa
Exposure time: 10s
Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
NLRP3 was immunoprecipitated from 0.35 mg RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab263899 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263899 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/1000 dilution.
Lane 1: RAW 264.7 whole cell lysate 10μg
Lane 2: ab263899 IP in RAW 264.7 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab263899 in RAW 264.7 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
All lanes: Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
Predicted band size: 118 kDa
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
The molecular weight/expression profile observed is consistent with what has been described in the literature (PMID: 26939933, 30315268).
Note: The band around 75kDa is the short form of NLRP3 (NLRP3s).
Low expression cell line: Jurkat (PMID: 19767079).
Exposure times: Lane 1: 48 secs; Lane 2: 3 min; Lanes 3-4: 1 sec; Lane 5: 6 secs.
All lanes: Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution
Lane 1: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg
Lane 2: Jurkat (human t cell leukemia t lymphocyte) whole cell lysate at 20 µg
Lane 3: Untreated RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 10 µg
Lane 4: RAW 264.7 treated with 10 μg/ml LPS for 8 hours, whole cell lysate at 10 µg
Lane 5: J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 118 kDa
The molecular weight/expression profile observed is consistent with what has been described in the literature (PMID: 26939933, 30315268).
Note: The band around 75kDa is the short form of NLRP3 (NLRP3s).
Low expression cell line: Jurkat (PMID: 19767079).
Exposure times: Lane 1: 48 secs; Lane 2: 3 min; Lanes 3-4: 1 sec; Lane 5: 6 secs.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263899).
Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
NLRP3 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate with ab263899 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263899 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/1000 dilution.
Lane 1: THP-1 whole cell lysate 10μg
Lane 2: ab263899 IP in THP-1 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab263899 in THP-1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
All lanes: Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
Predicted band size: 118 kDa
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
Stimulation is required to allow detection of the NLRP3 protein in C6, PC-12 and NIH/3T3 cell lines.
All lanes: Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution
Lane 1: C6 (Rat glial tumor glial cell) whole cell lysate at 10 µg
Lane 2: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 10 µg
Lane 3: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg
Lane 4: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Predicted band size: 118 kDa
Observed band size: 118 kDa
Exposure time: 15s
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
The molecular weight/expression profile observed is consistent with what has been described in the literature (PMID: 26939933, 30315268).
All lanes: Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution
All lanes: Rat spleen lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 118 kDa
Exposure time: 3min
The molecular weight/expression profile observed is consistent with what has been described in the literature (PMID: 26939933, 30315268).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263899).
Indirect ELISA - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
ELISA analysis of NLRP3 recombinant protein at 1000 ng/mL with ab263899. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
Blocking and Diluting buffer: 5% NFDM/TBST

Lane 9 Exposure Time: 20 seconds

NLRP3 is expressed at a much low level in normal tissues (PMID: 33538177, PMID: 25524927, PMID: 20688930, PMID: 35676979, PMID: 31923221, PMID: 24166187).
All lanes: Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution
Lane 1: Mouse thymus tissue lysate at 20 µg
Lane 2: Mouse lung tissue lysate at 20 µg
Lane 3: Mouse heart tissue lysate at 20 µg
Lane 4: Mouse kidney tissue lysate at 20 µg
Lane 5: Mouse brain tissue lysate at 20 µg
Lane 6: Mouse placenta tissue lysate at 20 µg
Lane 7: Mouse ovary tissue lysate at 20 µg
Lane 8: Mouse liver tissue lysate at 20 µg
Lane 9: Raw 264.7(Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 118 kDa
Observed band size: 118 kDa
Exposure time: 180s
Flow Cytometry (Intracellular) - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte) cells labeling NLRP3 with ab263899 at 1/50 (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
Blocking and dilution buffer: 5% NFDM /TBST. NLRP3 is expressed at a much low level in normal tissues (PMID: 33538177, PMID: 25524927, PMID: 20688930, PMID: 35676979, PMID: 31923221, PMID: 24166187).
All lanes: Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution
Lane 1: Rat spleen tissue lysate at 20 µg
Lane 2: Rat thymus tissue lysate at 20 µg
Lane 3: Rat lung tissue lysate at 20 µg
Lane 4: Rat heart tissue lysate at 20 µg
Lane 5: Rat kidney tissue lysate at 20 µg
Lane 6: Rat brain tissue lysate at 20 µg
Lane 7: Rat hippocampus tissue lysate at 20 µg
Lane 8: Rat spinal cord tissue lysate at 20 µg
Lane 9: Rat placenta tissue lysate at 20 µg
Lane 10: Rat ovary tissue lysate at 20 µg
Lane 11: Rat liver tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Exposure time: 180s
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)
ab263899 was shown to react with NLRP3 in wild-type THP-1 cells in Western blot with loss of signal observed in a NLRP3 knockout cell line. Wild-type THP-1 and NLRP3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab263899 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution
Lane 1: Wild-type THP-1 lysate at 20 µg
Lane 2: Wild-type THP-1 lysate treated with 200 ng/ml PMA, 48 hr at 20 µg
Lane 3: NLRP3 knock-out THP-1 lysate at 20 µg
Lane 4: NLRP3 knock-out THP-1 lysate treated with 200 ng/ml PMA, 48 hr at 20 µg