Anti-CIAS1/NALP3/NLRP3 Antibody

Product Name

Anti-CIAS1/NALP3/NLRP3 Antibody

View all NLRP3/NALP3 Antibodies

SKU/Catalog Number

PA1665

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CIAS1/NALP3/NLRP3 Antibody catalog # PA1665. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-CIAS1/NALP3/NLRP3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1665)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the N-terminus of human CIAS1, different from the related rat and mouse sequences by one amino acid.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PA1665 is reactive to NLRP3 in Human, Mouse, Rat

Applications

PA1665 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

110 kDa

Calculated molecular weight

118173 MW

Background of NLRP3/NALP3

NLRP3 (NLR FAMILY, PYRIN DOMAIN-CONTAINING 3), also known as CIAS1, CRYOPYRIN, NALP3 or PYPAF1, is a protein that in humans is encoded by the NLRP3 (NOD-like receptor family, pryin domain containing 3) gene. The NLRP3 gene encodes a pyrin-like protein expressed predominantly in peripheral blood leukocytes. And the NLRP3 gene is mapped on 1q44. NLRP3 interacts with apoptosis-associated speck-like protein containing a CARD (ASC). The encoded protein may play a role in the regulation of inflammation and apoptosis. Mutation of the NALP3 nucleotide-binding domain reduced ATP binding, CASP1 activation, IL1B production, cell death, macromolecular complex formation, self-association, and association with ASC. Consistent with an essential role for Nlrp3 inflammasomes in antifungal immunity, Gross et al.showed that Nlrp3-deficient mice are hypersusceptible to C. albicans infection. Activation of the NLRP3 inflammasome in response to virus or to RNA was dependent upon lysosomal maturation and reactive oxygen species production in human cells. The NLRP3 inflammasome senses obesity-associated danger signals and contributes to obesity-induced inflammation and insulin resistance.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconstitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, By Heat
Flow Cytometry(Fixed), 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of NLRP3 using anti-NLRP3 antibody (PA1665).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human THP-1 whole cell lysates,
Lane 2: rat PC-12 whole cell lysates,
Lane 3: mouse spleen whole cell lysates,
Lane 4: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NLRP3 antigen affinity purified polyclonal antibody (Catalog # PA1665) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for NLRP3 at approximately 110 kDa. The expected band size for NLRP3 is at 118 kDa.

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Figure 2. IHC analysis of NLRP3 using anti-NLRP3 antibody (PA1665).
NLRP3 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NLRP3 Antibody (PA1665) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of NLRP3 using anti-NLRP3 antibody (PA1665).
NLRP3 was detected in a paraffin-embedded section of human squamous cell cervical carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-NLRP3 Antibody (PA1665) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of THP-1 cells using anti-NLRP3 antibody (PA1665).
Overlay histogram showing THP-1 cells stained with PA1665 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NLRP3 Antibody (PA1665, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For NLRP3 (Source: Uniprot.org, NCBI)

Alternative Names

AGTAVPRL; AII; AII/AVP; Angiotensin/vasopressin receptor AII/AVP-like; mouse NLRP3; NLRP3; AVP; C1orf7; Caterpiller protein 1.1; CIAS1; CLR1.1; Cold autoinflammatory syndrome 1 protein; Cold autoinflammatory syndrome 1; Cryopyrin; Familial cold autoinflammatory syndrome; FCAS; FCU; Leucine-rich repeat-, and PYD-containing protein 3; Muckle-Wells syndrome; MWS; NACHT; NACHT, LRR and PYD containing protein 3; NACHT, LRR and PYD domains-containing protein 3; NALP3; NLR family, pyrin domain containing 3; NLRP3 mouse; NLRP3 polyclonal; NLRP3 rat; NLRP3; Nucleotide-Binding Oligomerization Domain, Le NLRP3 AGTAVPRL, AII, AVP, C1orf7, CIAS1, CLR1.1, DFNA34, FCAS, FCAS1, FCU, KEFH, MWS, NALP3, PYPAF1 NLR family pyrin domain containing 3 NACHT, LRR and PYD domains-containing protein 3|NACHT domain-, leucine-rich repeat-, and PYD-containing protein 3|NACHT, LRR and PYD containing protein 3|PYRIN-containing APAF1-like protein 1|caterpiller protein 1.1|cold autoinflammatory syndrome 1 protein|cold-induced autoinflammatory syndrome 1 protein|cryopyrin|cryopyrin, NACHT, LRR and PYD domains - containing protein 3|deafness, autosomal dominant 34|nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing 3

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