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NLRP3 Kit ELISA

NLRP3 Reactivité: Humain Colorimetric Sandwich ELISA 0.156 ng/mL - 10 ng/mL Plasma, Serum, Tissue Homogenate
N° du produit ABIN6973463
  • Antigène Voir toutes NLRP3 Kits ELISA
    NLRP3 (NLR Family, Pyrin Domain Containing 3 (NLRP3))
    Reactivité
    • 2
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    0.156 ng/mL - 10 ng/mL
    Seuil minimal de détection
    0.156 ng/mL
    Application
    ELISA
    Fonction
    For the quantitative determination of human NACHT, LRR and PYD domains-containing protein 3 (NLRP3/C1orf7/CIAS1/NALP3/PYPAF1) concentrations in serum, cell culture supernates, cerebrospinal fluid (CSF).
    Type d'échantillon
    Plasma, Serum, Tissue Homogenate
    Analytical Method
    Quantitative
    Specificité
    This assay has high sensitivity and excellent specificity for detection of human NLRP3. No significant cross-reactivity or interference between human NLRP3 and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between human NLRP3 and all the analogues, therefore, cross reaction may still exist.
    Sensibilité
    0.039 ng/mL
    Ingrédients
    • Assay plate
    • Standard
    • HRP-avidin (100 x concentrate)
    • Biotin-antibody (100 x concentrate)
    • Sample Diluent
    • HRP-avidin Diluent
    • Biotin-antibody Diluent
    • Wash Buffer (25 x concentrate)
    • TMB Substrate
    • Stop Solution
    • Adhesive Strip
    Top Product
    Discover our top product NLRP3 Kit ELISA
  • Indications d'application
    Optimal working dilution should be determined by the investigator.
    Volume d'échantillon
    100 μL
    Durée du test
    1 - 4.5 h
    Plaque
    Pre-coated
    Protocole
    1. Prepare reagents, samples and standards as instructed.
    2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
    3. Remove the liquid of each well, don't wash.
    4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
    5. Aspirate and wash 3 times.
    6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
    7. Aspirate and wash 5 times.
    8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
    9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.
    Préparation des réactifs
    1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
    2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
    3. Wash Buffer (1x) - If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
    4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 10 ng/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (10 ng/mL). Sample Diluent serves as the zero standard (0 ng/mL).
    Note:
    • Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
    • Bring all reagents to room temperature (18-25 °C) before use for 30 min.
    • Prepare fresh standard for each assay. Use within 4 hours and discard after use.
    • Making serial dilution in the wells directly is not permitted.
    • Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
    • Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.
    Préparation de l'échantillon
    • It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
    • If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
    • If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
    • Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).
    Précision du teste
    Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
    Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.
    Restrictions
    For Research Use only
  • Stock
    4 °C,-20 °C
    Stockage commentaire
    Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.
    Date de péremption
    6 months
  • Loukovaara, Piippo, Kinnunen, Hytti, Kaarniranta, Kauppinen: "NLRP3 inflammasome activation is associated with proliferative diabetic retinopathy." dans: Acta ophthalmologica, Vol. 95, Issue 8, pp. 803-808, (2018) (PubMed).

    Kinnunen, Piippo, Loukovaara, Hytti, Kaarniranta, Kauppinen: "Lysosomal destabilization activates the NLRP3 inflammasome in human umbilical vein endothelial cells (HUVECs)." dans: Journal of cell communication and signaling, Vol. 11, Issue 3, pp. 275-279, (2017) (PubMed).

    Piippo, Korkmaz, Hytti, Kinnunen, Salminen, Atalay, Kaarniranta, Kauppinen: "Decline in cellular clearance systems induces inflammasome signaling in human ARPE-19 cells." dans: Biochimica et biophysica acta, Vol. 1843, Issue 12, pp. 3038-46, (2014) (PubMed).

  • Antigène Voir toutes NLRP3 Kits ELISA
    NLRP3 (NLR Family, Pyrin Domain Containing 3 (NLRP3))
    Autre désignation
    NLR family, pyrin domain containing 3 (NLRP3 Produits)
    Synonymes
    CIAS1 Kit ELISA, Nalp3 Kit ELISA, avp Kit ELISA, cias1 Kit ELISA, nalp3 Kit ELISA, pypaf1 Kit ELISA, agtavprl Kit ELISA, AGTAVPRL Kit ELISA, AII Kit ELISA, AVP Kit ELISA, C1orf7 Kit ELISA, CLR1.1 Kit ELISA, FCAS Kit ELISA, FCU Kit ELISA, MWS Kit ELISA, NALP3 Kit ELISA, PYPAF1 Kit ELISA, AII/AVP Kit ELISA, Cias1 Kit ELISA, Mmig1 Kit ELISA, Pypaf1 Kit ELISA, NLR family pyrin domain containing-like Kit ELISA, NLR family pyrin domain containing 3 Kit ELISA, NLR family, pyrin domain containing 3 L homeolog Kit ELISA, NLR family, pyrin domain containing 3 Kit ELISA, NLRPL Kit ELISA, NLRP3 Kit ELISA, nlrp3.L Kit ELISA, Nlrp3 Kit ELISA
    Sujet

    Abbreviation: NLRP3

    Alias: AGTAVPRL, AII, AII/AVP, AVP, C1orf7, CIAS1, CLR1.1, FCAS, FCU, FLJ95925, MWS, NALP3, PYPAF1, AII/AVP receptor-like|NACHT domain-, leucine-rich repeat-, and PYD-containing protein 3|NACHT, LRR and PY

    UniProt
    Q96P20
    Pathways
    Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Endopeptidase Activity, Inflammasome
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