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Knockout Tested Rabbit Recombinant Multiclonal NLRP3 antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra), IP and reacts with Human, Mouse, Rat samples.

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Images

Flow Cytometry (Intracellular) - Anti-NLRP3 antibody [RM1021] (AB283819), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-NLRP3 antibody [RM1021] (AB283819), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-NLRP3 antibody [RM1021] (AB283819), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-NLRP3 antibody [RM1021] (AB283819), expandable thumbnail
  • Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (AB283819), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Multiclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IFFlow Cyt (Intra)IP
Human
Not recommended
Tested
Tested
Tested
Tested
Mouse
Not recommended
Tested
Tested
Tested
Tested
Rat
Not recommended
Tested
Expected
Expected
Expected

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Rat

Dilution info

1/1000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50

Notes

-

Species

Mouse

Dilution info

1/50

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50

Notes

-

Species

Mouse

Dilution info

1/500

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Species

Mouse

Dilution info

1/30

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

Function

As the sensor component of the NLRP3 inflammasome, plays a crucial role in innate immunity and inflammation. In response to pathogens and other damage-associated signals, initiates the formation of the inflammasome polymeric complex, made of NLRP3, PYCARD and CASP1 (and possibly CASP4 and CASP5). Recruitment of proCASP1 to the inflammasome promotes its activation and CASP1-catalyzed IL1B and IL18 maturation and secretion in the extracellular milieu (PubMed:28847925). Activation of NLRP3 inflammasome is also required for HMGB1 secretion (PubMed:22801494). The active cytokines and HMGB1 stimulate inflammatory responses. Inflammasomes can also induce pyroptosis, an inflammatory form of programmed cell death. Under resting conditions, NLRP3 is autoinhibited. NLRP3 activation stimuli include extracellular ATP, reactive oxygen species, K(+) efflux, crystals of monosodium urate or cholesterol, amyloid-beta fibers, environmental or industrial particles and nanoparticles, cytosolic dsRNA, etc. However, it is unclear what constitutes the direct NLRP3 activator. Activation in presence of cytosolic dsRNA is mediated by DHX33 (PubMed:23871209). 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. Binds to the consensus DNA sequence 5'-GRRGGNRGAG-3'. May also participate in the transcription of IL5, IL13, GATA3, CCR3, CCR4 and MAF (By similarity).

Targets

Nlrp3

Alternative names

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, C1orf7, CIAS1, NALP3, PYPAF1, NLRP3

Recommended products

Knockout Tested Rabbit Recombinant Multiclonal NLRP3 antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra), IP and reacts with Human, Mouse, Rat samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Multiclonal

Immunogens
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

RM1021

Purification technique

Affinity purification Protein A

Concentration
Loading...

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

This product is a recombinant multiclonal 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.

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

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.

8 product images

  • Flow Cytometry (Intracellular) - Anti-NLRP3 antibody [RM1021] (ab283819), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-NLRP3 antibody [RM1021] (ab283819)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte) treated with 50nM PMA and 5ug/ml LPS for 24h(Red) / Untreated control (Green) cells labelling NLRP3 with ab283819 at 1/50 dilution (1ug) (Red) and Green (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled 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) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

  • Flow Cytometry (Intracellular) - Anti-NLRP3 antibody [RM1021] (ab283819), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-NLRP3 antibody [RM1021] (ab283819)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 10ug/ml LPS for 8h(Red) / Untreated control (Green) cells labelling NLRP3 with ab283819 at 1/500 dilution (0.1ug) (Red) and Green (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled 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) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

  • Immunocytochemistry/ Immunofluorescence - Anti-NLRP3 antibody [RM1021] (ab283819), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-NLRP3 antibody [RM1021] (ab283819)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling NLRP3 with ab283819 at 1/50 (11.4 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing increased mainly cytoplasmic staining in THP-1 cells treated with 12-O-Tetradecanoylphorbol-13-acetate (80 nM) for 16 h then replaced it with lipopolysaccharide (1 ug/ml) for 8 h. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-NLRP3 antibody [RM1021] (ab283819), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-NLRP3 antibody [RM1021] (ab283819)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling NLRP3 with ab283819 at 1/50 (11.4 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing increased cytoplasmic staining in RAW 264.7 cells treated with lipopolysaccharide (10 ug/ml) for 8 h. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

  • Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (ab283819), expandable thumbnail

    Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (ab283819)

    NLRP3 was immunoprecipitated from 0.35 mg RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10ug with ab283819 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283819 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
    Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 µg
    Lane 2: ab283819 IP in RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab283819 in RAW 264.7 whole cell lysate
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Lower bands should be cleavage fragment of NLRP3. (PMID: 26119741)

    All lanes: Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (ab283819) at 1/30 dilution

    All lanes: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

    Secondary

    All lanes: Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (ab283819) at 1/5000 dilution

    Exposure time: 84s

  • Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (ab283819), expandable thumbnail

    Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (ab283819)

    NLRP3 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate 10ug with ab283819 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283819 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
    Lane 1: THP-1 (Human monocytic leukemia monocyte) whole cell lysate
    Lane 2: ab283819 IP in THP-1 (Human monocytic leukemia monocyte) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab283819 in THP-1 whole cell lysate
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Lower bands should be cleavage fragment of NLRP3. (PMID: 26119741)

    All lanes: Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (ab283819) at 1/30 dilution

    All lanes: THP-1 (Human monocytic leukemia monocyte) whole cell lysate

    Secondary

    All lanes: Immunoprecipitation - Anti-NLRP3 antibody [RM1021] (ab283819) at 1/5000 dilution

    Exposure time: 84s

  • Western blot - Anti-NLRP3 antibody [RM1021] (ab283819), expandable thumbnail

    Western blot - Anti-NLRP3 antibody [RM1021] (ab283819)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Negative control: Jurkat (PMID:19767079).

    Exposure time: Lane1-3: 3min; Lane4-5: 37 seconds; Lane6-7: 10 seconds

    All lanes: Western blot - Anti-NLRP3 antibody [RM1021] (ab283819) at 1/1000 dilution

    Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

    Lane 2: THP-1 (Human monocytic leukemia monocyte) whole cell lysate at 20 µg

    Lane 3: THP-1 treated with 80nM TPA overnight, then 1μg/ml lipopolysaccharide (LPS) for 8h whole cell lysate at 20 µg

    Lane 4: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

    Lane 5: RAW 264.7 treated with 1μg/ml lipopolysaccharide (LPS) for 6h whole cell lysate at 20 µg

    Lane 6: NR8383 (Rat lu macrophage (alveolar)) whole cell lysate at 20 µg

    Lane 7: NR8383 treated with 1μg/ml lipopolysaccharide (LPS) for 4h, then add 1μg/ml BFA for another 3h 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

    Observed band size: 118 kDa

  • Western blot - Anti-NLRP3 antibody [RM1021] (ab283819), expandable thumbnail

    Western blot - Anti-NLRP3 antibody [RM1021] (ab283819)

    Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS
    Lysates at 20 µg per lane.
    The samples were run on a Bis-Tris gel under reducing conditions.
    Western blot: Anti-NLRP3 antibody (ab283819) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red.
    In Western blot, ab283819 was shown to bind specifically to NLRP3. Target of interest was observed at 118 kDa in untreated wild-type THP-1 cell lysates (lane 1) 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) (lane 2). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

    All lanes: Western blot - Anti-NLRP3 antibody [RM1021] (ab283819) at 1/1000 dilution

    Lane 1: Wild-type THP-1 (Human monocytic leukemia monocyte) whole cell lysate at 20 µg

    Lane 2: NLRP3 knockout THP-1 whole cell lysate at 20 µg

    Secondary

    Lanes 1 - 2: Goat Anti-Rabbit IgG H&L (800CW) at 1/20000 dilution

    Lanes 1 - 2: Goat anti-Mouse IgG H&L (680RD) at 1/20000 dilution

    Observed band size: 118 kDa, 37 kDa

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