Human/Mouse NLRP3/NALP3 Antibody

(9 citations)
(2 Reviews)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects mouse NLRP3/NALP3 in direct ELISAs. In direct ELISAs, 100% cross-reactivity with recombinant human NLRP3/NALP3 is observed.
  • Source
    Monoclonal Rat IgG2A Clone # 768319
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant mouse NLRP3/NALP3
    Met1-Arg153
    Accession # Q8R4B8
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Mouse NLRP3/NALP3 by Western Blot. Western blot shows lysates of J774A.1 mouse reticulum cell sarcoma macrophage cell line and RAW 264.7 mouse monocyte/macrophage cell line. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human/Mouse NLRP3/NALP3 Monoclonal Antibody (Catalog # MAB7578) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for NLRP3/NALP3 at approximately 117 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
NLRP3/NALP3 in RAW 264.7 Mouse Cell Line. NLRP3/NALP3 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Rat Anti-Human/Mouse NLRP3/NALP3 Monoclonal Antibody (Catalog # MAB7578) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern­Lights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Intracellular Staining by Flow Cytometry
Detection of NLRP3/NALP3 in RAW 264.7 Mouse Cell Line by Flow Cytometry. RAW 264.7 mouse monocyte/macrophage cell line was stained with Rat Anti-Human/Mouse NLRP3/ NALP3 Monoclonal Antibody (Catalog # MAB7578, filled histo­gram) or isotype control antibody (Catalog # MAB006, open histo­gram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Intracellular Staining by Flow Cytometry
Detection of NLRP3/NALP3 in Human Monocytes by Flow Cytometry. Human peripheral blood monocytes was stained with Rat Anti-Human/Mouse NLRP3/NALP3 Monoclonal Antibody (Catalog # MAB7578, filled histogram) or isotype control antibody (Catalog # MAB006, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: NLRP3/NALP3
NLRP3, also known as NALP3, CIAS1, PYPAF or Cryopyrin, is a cytosolic ~120 kDa member of the NLRP family of proteins expressed in leukocytes, especially neutrophils. As a component of the inflammasome, NLRP3 activates caspases 1 and 5. Defects in NLRP3 may cause FCAS1, CINCA, or Muckle-Wells autoinflammatory syndromes. Mouse NLRP3 contains an N-terminal Pyrin domain (aa 1-91) followed by a Nacht region (aa 216-532), and seven LRRs (aa 739-988). Within the sequence used as an immunogen, mouse NLRP3 shares 78% and 93% aa identity with human and rat NLRP3, respectively. Alternate splicing of mouse NLRP3 generates additional isoforms that lack either LRR2 and 3, LRR 6 and 7, or LRR4-9.
  • Long Name:
    NLR Family, Pyrin Domain Containing 2
  • Entrez Gene IDs:
    114548 (Human); 216799 (Mouse); 287362 (Rat)
  • Alternate Names:
    AGTAVPRL; AII; AII/AVP; Angiotensin/vasopressin receptor AII/AVP-like; 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; Nucleotide-Binding Oligomerization Domain, Leucine Rich Repeat And Pyrin Domain Containing 3; PYPAF1; PYRIN-containing APAF1-like protein 1
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

9 Citations: Showing 1 - 9
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Applications
Sample Type
  1. Inhibition of NLRP3 Inflammasome Pathway by Butyrate Improves Corneal Wound Healing in Corneal Alkali Burn
    Authors: F Bian, Y Xiao, M Zaheer, EA Volpe, SC Pflugfelde, DQ Li, CS de Paiva
    Int J Mol Sci, 2017;18(3):.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC - Frozen
  2. The Antimalarial Chloroquine Suppresses LPS-Induced NLRP3 Inflammasome Activation and Confers Protection against Murine Endotoxic Shock
    Authors: X Chen, N Wang, Y Zhu, Y Lu, X Liu, J Zheng
    Mediators Inflamm., 2017;2017(0):6543237.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  3. Intestinal fungi contribute to development of alcoholic liver disease
    Authors: AM Yang, T Inamine, K Hochrath, P Chen, L Wang, C Llorente, S Bluemel, P Hartmann, J Xu, Y Koyama, T Kisseleva, MG Torralba, K Moncera, K Beeri, CS Chen, K Freese, C Hellerbran, SM Lee, HM Hoffman, WZ Mehal, G Garcia-Tsa, EA Mutlu, A Keshavarzi, GD Brown, SB Ho, R Bataller, P Stärkel, DE Fouts, B Schnabl
    J. Clin. Invest., 2017;0(0):.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  4. Knockout of Ccr2 alleviates photoreceptor cell death in rodent retina exposed to chronic blue light
    Cell Death Dis, 2016;7(11):e2468.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC - Frozen
  5. The cyclopentenone prostaglandin 15d-PGJ2 inhibits the NLRP1 and NLRP3 inflammasomes.
    Authors: Maier N, Leppla S, Moayeri M
    J Immunol, 2015;194(6):2776-85.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: IP
  6. Chinese Herbal Preparation Xuebijing Potently Inhibits Inflammasome Activation in Hepatocytes and Ameliorates Mouse Liver Ischemia-Reperfusion Injury.
    Authors: Liu X, Hu Z, Zhou B, Li X, Tao R
    PLoS ONE, 2015;10(7):e0131436.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  7. Suppression of NLRP3 inflammasome by gamma-tocotrienol ameliorates type 2 diabetes.
    Authors: Kim Y, Wang W, Okla M, Kang I, Moreau R, Chung S
    J Lipid Res, 2015;57(1):66-76.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  8. Caspase-8 as an Effector and Regulator of NLRP3 Inflammasome Signaling.
    Authors: Antonopoulos C, Russo H, El Sanadi C, Martin B, Li X, Kaiser W, Mocarski E, Dubyak G
    J Biol Chem, 2015;290(33):20167-84.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  9. ASC in renal collecting duct epithelial cells contributes to inflammation and injury after unilateral ureteral obstruction.
    Authors: Komada T, Usui F, Shirasuna K, Kawashima A, Kimura H, Karasawa T, Nishimura S, Sagara J, Noda T, Taniguchi S, Muto S, Nagata D, Kusano E, Takahashi M
    Am J Pathol, 2014;184(5):1287-98.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
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Average Rating: 4 (Based on 2 reviews)

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We have 2 reviews tested in 1 species: Mouse.
We have 2 reviews tested in 2 applications: Immunocytochemistry/Immunofluorescence, Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Immunocytochemistry/Immunofluorescence Human/Mouse NLRP3/NALP3 Antibody MAB7578
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Excellent
 ICC/IF Mouse Anonymous 07/21/2017
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Immunocytochemistry/Immunofluorescence Human/Mouse NLRP3/NALP3 Antibody MAB7578
Immunocytochemistry/Immunofluorescence: Human/Mouse NLRP3/NALP3 Antibody [MAB7578]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedJ774A.1 mouse reticulum cell sarcoma macrophage cell line
SpeciesMouse
Western Blot Human/Mouse NLRP3/NALP3 Antibody MAB7578
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Average
 WB Mouse Anonymous 01/11/2017
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Western Blot Human/Mouse NLRP3/NALP3 Antibody MAB7578
Western Blot: Human/Mouse NLRP3/NALP3 Antibody [MAB7578]

Summary

ApplicationWestern Blot
Sample Testedprimary macrophages
SpeciesMouse

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