Anti-CIAS1/NALP3/NLRP3 Antibody

SKU PA1665
Size 100μg/vial
Reactivity Human, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications Flow Cytometry, IF, IHC-P, ICC, WB

Overview

Product Name Anti-CIAS1/NALP3/NLRP3 Antibody
SKU/Catalog Number PA1665
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for NACHT, LRR and PYD domains-containing protein 3(NLRP3) detection. Tested with WB, IHC-P, ICC/IF, FCM in Human;Mouse;Rat.
Cite This Product Anti-CIAS1/NALP3/NLRP3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1665)
Host Rabbit
Contents/Buffer Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence at the N-terminus of human CIAS1(12-31aa RYLEDLEDVDLKKFKMHLED), different from the related rat and mouse sequences by one amino acid.
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, Mouse, By Heat
Western blot, 0.1-0.5μg/ml, Human
Immunocytochemistry/ Immunofluorescence, 2μg/ml, Human, Rat
Flow Cytometry, 1-3μg/1x106 cells, Human

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P) and ICC.

Images And Assay Conditions

Figure 1. Western blot analysis of -CIAS1/NALP3 using anti- -CIAS1/NALP3 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 50ug of sample under reducing conditions.
Lane 1: HEP-2 Cell Lysate
Lane 2: A549 Cell Lysate
Lane 3: U87 Cell Lysate
Lane 4: CEM Cell Lysate
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- -CIAS1/NALP3 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 1:10000 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 -CIAS1/NALP3 at approximately 118KD. The expected band size for -CIAS1/NALP3 is at 118KD.

Figure 2. IHC analysis of CIAS1/NALP3 using anti- CIAS1/NALP3 antibody (PA1665).
CIAS1/NALP3 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- CIAS1/NALP3 Antibody (PA1665) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 3. Flow Cytometry analysis of THP-1 cells using anti-CIAS1/NALP3 antibody (PA1665).
Overlay histogram showing THP-1 cells stained with PA1665 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CIAS1/NALP3 Antibody (PA1665,1μg/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 (Red line) was also used as a control.

Figure 4. Flow Cytometry analysis of U937 cells using anti-CIAS1/NALP3 antibody (PA1665).
Overlay histogram showing U937 cells stained with PA1665 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CIAS1/NALP3 Antibody (PA1665,1μg/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 (Red line) was also used as a control.

Figure 5. IF analysis of CIAS1/NALP3 using anti- CIAS1/NALP3 antibody (PA1665).
CIAS1/NALP3 was detected in immunocytochemical section of THP-1 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti- CIAS1/NALP3 Antibody (PA1665) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 6. IF analysis of CIAS1/NALP3 using anti- CIAS1/NALP3 antibody (PA1665).
CIAS1/NALP3 was detected in immunocytochemical section of THP-1 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti- CIAS1/NALP3 Antibody (PA1665) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 9. IF analysis of CIAS1/NALP3 using anti- CIAS1/NALP3 antibody (PA1665)
CIAS1/NALP3 was detected in paraffin-embedded section of rat colon tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- CIAS1/NALP3 Antibody (PA1665) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 7. IF analysis of CIAS1/NALP3 using anti- CIAS1/NALP3 antibody (PA1665)
CIAS1/NALP3 was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- CIAS1/NALP3 Antibody (PA1665) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 8. IF analysis of CIAS1/NALP3 using anti- CIAS1/NALP3 antibody (PA1665)
CIAS1/NALP3 was detected in paraffin-embedded section of rat spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- CIAS1/NALP3 Antibody (PA1665) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 10. IF analysis of CIAS1/NALP3 using anti- CIAS1/NALP3 antibody (PA1665)
CIAS1/NALP3 was detected in paraffin-embedded section of rat colon tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti- CIAS1/NALP3 Antibody (PA1665) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 11. IHC analysis of NLRP3 using anti- NLRP3 antibody (PA1665).
NLRP3 was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- NLRP3 Antibody (PA1665) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id Q96P20
Gene Name NLRP3
Protein Name NACHT, LRR and PYD domains-containing protein 3
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. .
Alternative Names NACHT, LRR and PYD domains-containing protein 3;Angiotensin/vasopressin receptor AII/AVP-like;Caterpiller protein 1.1;CLR1.1;Cold autoinflammatory syndrome 1 protein;Cryopyrin;PYRIN-containing APAF1-like protein 1;NLRP3;C1orf7, CIAS1, NALP3, PYPAF1;
Subcellular Localization Cytoplasm .
Molecular Weight 118173 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein 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. .
Research Areas Human, Mouse, Rat

*You can search these to find other products in these research areas.
Background 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.

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Polyclonal antibody for NALP3/NLRP3 detection. Host: Rabbit.Size: 100μg/vial. Tested applications: IHC-P. Reactive species: Human. NALP3/NLRP3 information: Molecular Weight: 118173 MW; Subcellular Localization: Cytoplasm ; 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.
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In stock
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PA1665
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$280.00

Troubleshooting

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Publications

Wang B, Zhang F, Zhang H, Wang Z, Ma YN, Zhu MJ, Du M. Oncotarget. 2017 Nov 1;8(59):100411-100420. doi: 10.18632/oncotarget.22243. eCollection 2017 Nov 21. Alcohol intake aggravates adipose browning and muscle atrophy in cancer-associated cachexia
Xue Y, Wang H, Du M, Zhu Mj. J Nutr Biochem. 2014 Jul;25(7):758-64. Doi: 10.1016/J.Jnutbio.2014.03.009. Epub 2014 Apr 1. Maternal Obesity Induces Gut Inflammation And Impairs Gut Epithelial Barrier Function In Nonobese Diabetic Mice.
Wang B, Yang G, Liang X, Zhu M, Du M. Bmc Complement Altern Med. 2014 May 20;14:162. Doi: 10.1186/1472-6882-14-162. Grape Seed Extract Prevents Skeletal Muscle Wasting In Interleukin 10 Knockout Mice.

Customer Q&As

  • Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
    A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
  • Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
    A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
  • Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
    A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
  • Q: What should I use for negative control?
    A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
  • Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
    A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
  • Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
    A: You can find the immunogen sequence under "
  • Q: What is the expected band size? Why is it different than the observed band size?
    A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
  • Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
    A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
  • Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
    A: Check our protocols under the tech support tab.
  • Q: What are some alternative names that could be used to describe this product?
    A: Some common names include but are not limited to nlrp3 antibody, nalp3 antibody
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