Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all NLRP3/NALP3 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human NLRP3/NALP3
Ala4-Val223
Accession # Q96P20

Specificity

Detects human NLRP3/NALP3 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) NALP1 and rhNALP2 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human NLRP3/NALP3 (aa 4‑223) Antibody

Detection of Human NLRP3/NALP3 antibody by Western Blot.

Detection of Human NLRP3/NALP3 by Western Blot.

Western blot shows lysates of HEK293 human embryonic kidney cell line either mock transfected, transfected with human NALP1 or transfected with NLRP3/NALP3 and THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 20 ng/mL PMA for 72 hours and 1 µg/mL LPS for 24 hours. PVDF Membrane was probed with 0.5 µg/mL of Sheep Anti-Human NLRP3/NALP3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6789) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for NLRP3/NALP3 at approximately 115 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
NLRP3/NALP3 antibody in THP-1 Human Cell Line by Immunocytochemistry (ICC).

NLRP3/NALP3 in THP‑1 Human Cell Line.

NLRP3/NALP3 was detected in immersion fixed THP-1 human acute monocytic leukemia cells, unstimulated (lower panel) and stimulated (upper panel) with PMA and LPS, using Sheep Anti-Human NLRP3/NALP3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6789) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Human NLRP3/NALP3 antibody by Simple WesternTM.

Detection of Human NLRP3/NALP3 by Simple WesternTM.

Simple Western lane view shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with PMA and LPS, loaded at 0.2 mg/mL. A specific band was detected for NLRP3/NALP3 at approximately 111 kDa (as indicated) using 50 µg/mL of Sheep Anti-Human NLRP3/NALP3 (aa 4-223) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6789) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Human NLRP3/NALP3 by Western Blot

Detection of Human NLRP3/NALP3 by Western Blot

NLRP3s expression in Monocyte by Western blot: NLRP3s protein expression assessed by western blotting in monocyte of individuals with a diagnosis of either severe Alzheimer’s disease (AD), moderate Alzheimer’s disease (MILD) or Mild Cognitive Impairment (MCI) and of age- and sex-matched Healthy Controls (HC). The same protein concentration of whole-cells lysates was loaded into the gel, as confirmed by actin. Representative results obtained in un-stimulated or in LPS-primed and A beta 42-stimulated monocytes are presented in the upper panel. Quantitative evaluation (arbitrary unit, AU) of NLRP3s expression obtained comparing band density (normalized to actin) in un-stimulated or in LPS-primed A beta 42-stimulated monocytes is shown in the lower panel. (N = 5, mean ± s.e.m., T test, *p <0.05). (N = 5/group, mean ± S.E.M and statistical significance are shown. *(p <0.05) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26939933), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human NLRP3/NALP3 (aa 4‑223) Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed THP-1 human acute monocytic leukemia cell line

Simple Western

50 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line treated with PMA and LPS

Western Blot

0.5 µg/mL
Sample: HEK293 human embryonic kidney cell line either transfected with human NALP1 or transfected with NALP3 and THP‑1 human acute monocytic leukemia cell line treated with PMA and LPS

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: NLRP3/NALP3

NALP3 (NAcht, Leucine-rich repeat and PYD domain containing protein 3; also PYPAF1, CIAS1 and CLR1.1) is a cytoplasmic 115-130 kDa member of the NLRP family of molecules. It is selectively expressed, being found in stratified squamous and transitional epithelium, Hassell's corpuscles, neutrophils, dendritic cells and T cells. NALP3 promotes IL-1 beta and IL-18 maturation by activating caspase-1. It does so by forming a NALP3 inflammasome comprised of NALP3, ASC and caspase-1. Bacterial RNA and cell wall peptidoglycan can bind to NALP3, promoting (potentially) ATP binding, NALP3 oligomerization, and caspase activation. Human NALP3 is 1036 amino acids (aa) in length. It contains an N-terminal DAPIN domain (aa 1-93), a NACHT domain (aa 220-536), and seven consecutive LRRs (aa 740-991). Alternate splice forms are reported. Either individually, or in combination, there can be deletions of aa 836-892 or 721-777, and a premature truncation after Gly719 that generates a 70 kDa isoform. Over aa 3-223, human NALP3 shares 76% aa identity with mouse NALP3.

Long Name

NLR Family, Pyrin Domain Containing 2

Alternate Names

AGTAVPRL, AII, CIAS1, Cryopyrin, FCAS, FCU, MWS, NACHT, NALP3, PYPAF1

Entrez Gene IDs

114548 (Human); 216799 (Mouse); 287362 (Rat)

Gene Symbol

NLRP3

UniProt

Q96P20 (Human)

Additional NLRP3/NALP3 Products

Product Documents for Human NLRP3/NALP3 (aa 4‑223) Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human NLRP3/NALP3 (aa 4‑223) Antibody

For research use only

Citations for Human NLRP3/NALP3 (aa 4‑223) Antibody

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Human NLRP3/NALP3 (aa 4‑223) Antibody

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  • Q: Does this NLRP3/NALP3 antibodies come in lyophilized form?

    A: Yes, we carry this NLRP2 antibodies in lyophilized format.

  • Q: What is the immunogen sequence of this NLRP3/NALP3 antibodies antibody?

    A: An E. coli derived sequence corresponding to Ala4 - Val223 of human NLRP3/NALP3.

  • Q: Does this NLRP3/NALP3 antibodies come in lyophilized form?

    A: Yes, we carry this NLRP2 antibodies in lyophilized format.

  • Q: What is the immunogen sequence of this NLRP3/NALP3 antibodies antibody?

    A: An E. coli derived sequence corresponding to Ala4 - Val223 of human NLRP3/NALP3.

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