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Product Info Summary
| SKU: | A00034-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-CIAS1/NALP3/NLRP3 Antibody Picoband®
SKU/Catalog Number
A00034-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-CIAS1/NALP3/NLRP3 Antibody Picoband® catalog # A00034-2. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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 Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00034-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human CIAS1/NALP3/NLRP3 recombinant protein (Position: D812-S1035).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00034-2 is reactive to NLRP3 in Human, Mouse, Rat
Observed Molecular Weight
118 kDa
Calculated molecular weight
118.2 kDa
Background of NLRP3
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.
Application & Images
Applications
A00034-2 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee
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, 0.25-0.5μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Hela whole cell, human U-87MG whole cell, rat spleen tissue, rat PC-12 whole cell, mouse thymus tissue, mouse lung tissue, mouse spleen tissue
FCM: THP-1 cell
Validation Images & Assay Conditions
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Western blot analysis of NLRP3 using anti-NLRP3 antibody (A00034-2).
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 30ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human U-87MG whole cell lysates,
Lane 3: rat spleen tissue lysates,
Lane 4: rat PC-12 whole cell lysates,
Lane 5: mouse thymus tissue lysates,
Lane 6: mouse lung tissue lysates,
Lane 7: mouse spleen tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 # A00034-2) 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 118KD. The expected band size for NLRP3 is at 118KD.
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Flow Cytometry analysis of THP-1 cells using anti-NLRP3 antibody (A00034-2).
Overlay histogram showing THP-1 cells stained with A00034-2 (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 (A00034-2, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control."
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Effects of magnolol on NLRP3 inflammasome, caspase-1 and caspase-3 signaling pathway in ALD mice. Magnolol was given to mice for three times, and then alcohol was gavaged. The level of NLRP3 inflammasome, caspase-1 and caspase-3 was detected by western blotting analysis with the compared with the internal control (GAPDH). The data is presented as mean ± SD. (*P < 0.05, **P < 0.01, ***P < 0.001).
Index in PubMed under a CC BY license. PMID: 31920652
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NLRP3, IL-1β, and IL-18 expression levels in the LT among the three groups of LDL-R −/− mice fed the different diets. (a) WB to detect the NLRP3, IL-1β, and IL-18 expression levels in the LT. The blots edge was imprinted as a marker and were cut prior to hybridization with antibodies. (b) Bar graph depicts the quantification of NLRP3, IL-1β, and IL-18 expression levels. Data are expressed as mean ± SEM; n = 3 in each group. *P < 0.05, **P < 0.01 vs. LDL-R −/− HD group
Index in PubMed under a CC BY license. PMID: 37568105
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Gene expression of pyroptosis in the LTs obtained from the three groups of LDL-R −/− mice fed the different diets. Relative mRNA expression of NLRP3, IL-1β, and IL-18 in the LTs. Data are represented as mean ± SEM; n = 3 in each group. *P < 0.05, **P < 0.01 vs. LDL-R −/− HD group
Index in PubMed under a CC BY license. PMID: 37568105
Specific Publications For Anti-CIAS1/NALP3/NLRP3 Antibody Picoband® (A00034-2)
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