Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NLRP3 antibody [EPR23073-96] (ab270449)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23073-96] to NLRP3
  • Suitable for: IP, ICC, WB
  • Knockout validated
  • Reacts with: Mouse

Overview

  • Product name

    Anti-NLRP3 antibody [EPR23073-96]
    See all NLRP3 primary antibodies

  • Description

    Rabbit monoclonal [EPR23073-96] to NLRP3

  • Host species

    Rabbit

  • Specificity

    Stimulation may be required to allow detection of the NLRP3 protein in most samples.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Mouse
    ICC
    Mouse
    IP
    Mouse
    WB
    Mouse
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: RAW 264.7 (+/- treatment with LPS) and J774A.1 whole cell lysates. ICC: RAW 264.7 (+ treatment with LPS) cells. IP: RAW 264.7 (+ treatment with LPS) whole cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-NLRP3 antibody [EPR23073-96] (ab270449)
    Western blot - Anti-NLRP3 antibody [EPR23073-96] (ab270449) This image is courtesy of Benedikt Saller, from Prof. Dr. Olaf Groß’ lab at the University of Freiburg
    All lanes : Anti-NLRP3 antibody [EPR23073-96] (ab270449) at 1/1000 dilution

    Lane 1 : Wild-type LPS treated (3 hour, 50 ng/mL) Mouse BMDCs
    Lane 2 : NLRP3 knockout LPS treated (3 hour, 50 ng/mL) Mouse BMDCs

    Lysates/proteins at 25000 cells per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (H+L) (HRP) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 118 kDa


    Exposure time: 2 minutes

    See Abreview

  • Immunocytochemistry - Anti-NLRP3 antibody [EPR23073-96] (ab270449)
    Immunocytochemistry - Anti-NLRP3 antibody [EPR23073-96] (ab270449)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (untreated)(-) and  RAW 264.7 (+LPS 10ug/ml 8h)(+) cells labelling NLRP3 with ab270449 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 after treatment with LPS. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/ml) dilution.

  • Immunoprecipitation - Anti-NLRP3 antibody [EPR23073-96] (ab270449)
    Immunoprecipitation - Anti-NLRP3 antibody [EPR23073-96] (ab270449)

    NLRP3 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) (treated with 10 ug/ml Lipopolysaccharides (LPS) for 8 hours) whole cell lysate with ab270449 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270449 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: RAW 264.7 treated with 10 ug/ml Lipopolysaccharides (LPS) for 8 hours, whole cell lysate 10 ug

    Lane 2: ab270449 IP in RAW 264.7 treated with 10 ug/ml Lipopolysaccharides(LPS) for 8 hours, whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab270449 in RAW 264.7 treated with 10 ug/ml Lipopolysaccharides(LPS) for 8 hours, whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 24 seconds.

    The band around 75kDa corresponds to the short form of NLRP3.

  • Western blot - Anti-NLRP3 antibody [EPR23073-96] (ab270449)
    Western blot - Anti-NLRP3 antibody [EPR23073-96] (ab270449)
    All lanes : Anti-NLRP3 antibody [EPR23073-96] (ab270449) at 1/1000 dilution

    Lane 1 : Untreated RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
    Lane 2 : RAW 264.7 treated with 10 µg/ml Lipopolysaccharides (LPS) for 8 hours, whole cell lysate
    Lane 3 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
    Lanes 2-3 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051 at 1/100000 dilution

    Predicted band size: 118 kDa
    Observed band size: 118 kDa



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The molecular weight/expression profile observed is consistent with what has been described in the literature (PMID:26939933, 30315268 ).

    The band around 75kDa corresponds to the short form of NLRP3.

    Exposure time: 54 seconds. 

  • Anti-NLRP3 antibody [EPR23073-96] (ab270449)
    Anti-NLRP3 antibody [EPR23073-96] (ab270449)

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