Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23094-1] to NLRP3 - BSA and Azide free
  • Suitable for: ELISA, WB, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free
    See all NLRP3 primary antibodies

  • Description

    Rabbit monoclonal [EPR23094-1] to NLRP3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ELISA, WB, Flow Cyt, IPmore details
    Unsuitable for: ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: THP-1 and J774A.1 whole cell lysate. LPS treated RAW 264.7 whole cell lysate. Rat spleen lysate. Mouse Wt BMDCs treated with 50 ng/ml LPS, 3h; Human THP1 Wt cells treated with 200 ng/ml LPS, 3h. RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate. IP: RAW 264.7 and THP-1 whole cell lysate. Flow Cyt: THP-1 cells.

  • General notes

    ab264468 is the carrier-free version of ab263899. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Western blot - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

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


    Exposure time: 15 seconds
  • Western blot - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Western blot - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468) This image is courtesy of Benedikt Saller, from Prof. Dr. Olaf Groß’ lab at the University of Freiburg.
    All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution

    Lane 1 : Wild-type mouse BMDCs treated with 50 ng/ml LPS
    Lane 2 : NLRP3 knockout mouse BMDCs treated with 50 ng/ml LPS
    Lane 3 : Wild-type human THP1 cells treated with 200 ng/ml LPS

    Lysates/proteins at 25000 cells per lane.

    Secondary
    All lanes : Goat anti-Rabbit HRP at 1/5000 dilution

    Developed using the ECL technique.

    Predicted band size: 118 kDa


    Exposure time: 10 seconds


    Blocking step: 2% SMP in PBS/T for 60 minutes at RT.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263899).

  • Western blot - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Western blot - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    All lanes : Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution

    Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
    Lane 2 : Jurkat (human t cell leukemia t lymphocyte) whole cell lysate
    Lane 3 : Untreated RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 4 : RAW 264.7 treated with 10 µg/ml LPS for 8 hours, whole cell lysate
    Lane 5 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 118 kDa



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

    Note: The band around 75kDa is the short form of NLRP3 (NLRP3s).

    Low expression cell line: Jurkat (PMID: 19767079).

    Exposure times: Lane 1: 48 secs; Lane 2: 3 min; Lanes 3-4: 1 sec; Lane 5: 6 secs.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263899).

  • Western blot - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Western blot - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Anti-NLRP3 antibody [EPR23094-1] (ab263899) at 1/1000 dilution + Rat spleen lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 118 kDa


    Exposure time: 3 minutes


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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263899).

  • Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)

    NLRP3 was immunoprecipitated from 0.35 mg RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10µg with ab263899 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263899 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

    Lane 1: RAW 264.7 whole cell lysate 10µg

    Lane 2: ab263899 IP in RAW 264.7 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263899 in RAW 264.7 whole cell lysate

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

    Exposure time: 10 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263899).

  • Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Immunoprecipitation - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)

    NLRP3 was immunoprecipitated from 0.35 mg THP-1 (Human monocytic leukemia monocyte) whole cell lysate 10µg with ab263899 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263899 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

    Lane 1: THP-1 whole cell lysate 10µg

    Lane 2: ab263899 IP in THP-1 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263899 in THP-1 whole cell lysate

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

    Exposure time: 30 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263899).

  • Flow Cytometry - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Flow Cytometry - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte) cells labeling NLRP3 with ab263899 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263899).

  • ELISA - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    ELISA - Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)

    This data was developed using ab263899, the same antibody clone in a different buffer formulation.

    ELISA analysis of NLRP3 recombinant protein at 1000 ng/mL with ab263899. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
  • Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)
    Anti-NLRP3 antibody [EPR23094-1] - BSA and Azide free (ab264468)

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