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

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Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23094-1] to NLRP3
Suitable for: ELISA, WB, IP, Flow Cyt (Intra)
Knockout validated
Reacts with: Mouse, Rat, Human, Recombinant fragment

Carrier Free

Product name

Anti-NLRP3 antibody [EPR23094-1]
See all NLRP3 primary antibodies
Description

Rabbit monoclonal [EPR23094-1] to NLRP3
Host species

Rabbit
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).
Tested applications

Suitable for: ELISA, WB, IP, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF or IHC-P
Species reactivity

Reacts with: Mouse, Rat, Human, Recombinant fragment
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: THP-1 and J774A.1 whole cell lysate. LPS treated RAW 264.7 whole cell lysate. Rat spleen lysate. Mouse Wt BMDCs treated with 50 ng/ml LPS, 3h; Human THP1 Wt cells treated with 200 ng/ml LPS, 3h. RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate. IP: RAW 264.7 and THP-1 whole cell lysate. Flow Cyt (intra): THP-1 cells.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23094-1
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
- Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab263899 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
ELISA		Use at an assay dependent concentration.
WB	(2)	1/1000. Predicted molecular weight: 118 kDa. 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.
IP		1/30.
Flow Cyt (Intra)		1/50.

Notes
ELISA Use at an assay dependent concentration.
WB 1/1000. Predicted molecular weight: 118 kDa. 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.
IP 1/30.
Flow Cyt (Intra) 1/50.

Application notes

Is unsuitable for ICC/IF or IHC-P.

Function

May function as an inducer of apoptosis. Interacts selectively with ASC and this complex may function as an upstream activator of NF-kappa-B signaling. Inhibits TNF-alpha induced activation and nuclear translocation of RELA/NF-KB p65. Also inhibits transcriptional activity of RELA. Activates caspase-1 in response to a number of triggers including bacterial or viral infection which leads to processing and release of IL1B and IL18.
Tissue specificity

Expressed in blood leukocytes. Strongly expressed in polymorphonuclear cells and osteoblasts. Undetectable or expressed at a lower magnitude in B- and T-lymphoblasts, respectively. High level of expression detected in chondrocytes. Detected in non-keratinizing epithelia of oropharynx, esophagus and ectocervix and in the urothelial layer of the bladder.
Involvement in disease

Defects in NLRP3 are the cause of familial cold autoinflammatory syndrome type 1 (FCAS1) [MIM:120100]; also known as familial cold urticaria. FCAS are rare autosomal dominant systemic inflammatory diseases characterized by episodes of rash, arthralgia, fever and conjunctivitis after generalized exposure to cold.
Defects in NLRP3 are a cause of Muckle-Wells syndrome (MWS) [MIM:191900]; also known as urticaria-deafness-amyloidosis syndrome. MWS is a hereditary periodic fever syndrome characterized by fever, chronic recurrent urticaria, arthralgias, progressive sensorineural deafness, and reactive renal amyloidosis. The disease may be severe if generalized amyloidosis occurs.
Defects in NLRP3 are the cause of chronic infantile neurologic cutaneous and articular syndrome (CINCA) [MIM:607115]; also known as neonatal onset multisystem inflammatory disease (NOMID). CINCA is a rare congenital inflammatory disorder characterized by a triad of neonatal onset of cutaneous symptoms, chronic meningitis and joint manifestations with recurrent fever and inflammation.
Sequence similarities

Belongs to the NLRP family.
Contains 1 DAPIN domain.
Contains 9 LRR (leucine-rich) repeats.
Contains 1 NACHT domain.
Cellular localization

Cytoplasm.
Target information above from: UniProt accession Q96P20 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 114548 Human
- Entrez Gene: 216799 Mouse
- Entrez Gene: 287362 Rat
- Omim: 606416 Human
- SwissProt: Q96P20 Human
- SwissProt: Q8R4B8 Mouse
- Unigene: 159483 Human
- Unigene: 54174 Mouse
Alternative names
- AGTAVPRL antibody
- AII/AVP antibody
- Angiotensin/vasopressin receptor AII/AVP like antibody
- Angiotensin/vasopressin receptor AII/AVP-like antibody
- C1orf7 antibody
- Caterpiller protein 1.1 antibody
- CIAS 1 antibody
- CIAS1 antibody
- CLR1.1 antibody
- Cold autoinflammatory syndrome 1 antibody
- Cold autoinflammatory syndrome 1 protein antibody
- Cryopyrin antibody
- Familial cold autoinflammatory syndrome antibody
- FCAS antibody
- FCU antibody
- LRR and PYD domains-containing protein 3 antibody
- Muckle-Wells syndrome antibody
- MWS antibody
- NACHT antibody
- NACHT LRR and PYD containing protein 3 antibody
- NALP 3 antibody
- NALP3 antibody
- NALP3_HUMAN antibody
- NLR family pyrin domain containing 3 antibody
- NLRP3 antibody
- PYPAF 1 antibody
- PYPAF1 antibody
- PYRIN containing APAF1 like protein 1 antibody
- PYRIN-containing APAF1-like protein 1 antibody
see all

Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/500 dilution

Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : NLRP3 knockout THP-1 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 118 kDa
Observed band size: 118 kDa

False colour image of Western blot: Anti-NLRP3 antibody [EPR23094-1] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (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 ab280063 (knockout 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) at 1/20000 dilution.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)This image is courtesy of Benedikt Saller, from Prof. Dr. Olaf Groß’ lab at the University of Freiburg.

All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution

Lane 1 : Mouse Wt BMDCs treated with 50 ng/ml LPS, 3h.
Lane 2 : Mouse NLRP3-/- BMDCs treated with 50 ng/ml LPS, 3h.
Lane 3 : Human THP1 Wt cells treated with 200 ng/ml LPS, 3h.

Lysates/proteins at 25000 cells per lane.

Secondary
All lanes : Goat anti-Rabbit HRP at 1/5000 dilution

Developed using the ECL technique.

Predicted band size: 118 kDa

Exposure time: 10 seconds

Blocking step: 2% SMP in PBS/T for 60 minutes at RT.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

All lanes : 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 : 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.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 118 kDa
Observed band size: 118 kDa

Exposure time: 15 seconds

Stimulation is required to allow detection of the NLRP3 protein in C6, PC-12 and NIH/3T3 cell lines.
Western blot - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution + Rat spleen lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 118 kDa

Exposure time: 3 minutes

The molecular weight/expression profile observed is consistent with what has been described in the literature (PMID: 26939933, 30315268).
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 10µg 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) (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 (ab172730) instead of ab263899 in RAW 264.7 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.
Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] (ab263899)

NLRP3 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate 10µg 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) (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 (ab172730) instead of ab263899 in THP-1 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 30 seconds.
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 (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, ab150077) at 1/2000 dilution was used as the secondary antibody.
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.
Anti-NLRP3 antibody [EPR23094-1] (ab263899)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab263899? Please let us know so that we can cite the reference in this datasheet.

ab263899 has been referenced in 49 publications.

Cai L et al. Glucocorticoid receptors involved in melatonin inhibiting cell apoptosis and NLRP3 inflammasome activation caused by bacterial toxin pyocyanin in colon. Free Radic Biol Med 162:478-489 (2021). WB ; Human . PubMed: 33189867
Yang Y et al. Impairment of sirtuin 1-mediated DNA repair is involved in bisphenol A-induced aggravation of macrophage inflammation and atherosclerosis. Chemosphere 265:128997 (2021). WB ; Mouse . PubMed: 33239236
Guo H et al. Carthamin yellow improves cerebral ischemia-reperfusion injury by attenuating inflammation and ferroptosis in rats. Int J Mol Med 47:1 (2021). WB ; Rat . PubMed: 33576458
Li Z et al. Long noncoding RNA MEG3 contributes to dysfunction of brain microvascular endothelial cells after intracerebral hemorrhage by regulating the miR-1930-5p/Mllt1 axis. Brain Res Bull 166:1-11 (2021). WB ; Mouse . PubMed: 33127454
Chai GR et al. Quercetin protects against diabetic retinopathy in rats by inducing heme oxygenase-1 expression. Neural Regen Res 16:1344-1350 (2021). WB ; Rat . PubMed: 33318415

View all Publications for this product

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1-2 of 2 Abreviews or Q&A

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Lysates of BMDCs and Thp1)

Gel Running Conditions

Reduced Denaturing (Continous 12% gel)

Loading amount

25000 cells

Treatment

LPS (3h, 50 ng/ml for BMDCs or 200 ng/ml for Thp1)

Specification

Lysates of BMDCs and Thp1

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Benedikt Saller

Verified customer

Submitted Jul 02 2020

Application

Western blot

Sample

Rat Cell lysate - whole cell (endothelial cells)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

30 µg

Specification

endothelial cells

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Dhanush Haspula

Verified customer

Submitted May 26 2020

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (49)

Customer reviews and Q&As

Western blot abreview for Anti-NLRP3 antibody [EPR23094-1]

Western blot abreview for Anti-NLRP3 antibody [EPR23094-1]