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NF-kB p65 Kit ELISA

NFkBP65 Reactivité: Humain Colorimetric Sandwich ELISA 0.16 ng/mL - 10 ng/mL Cell Culture Supernatant, Plasma, Serum
N° du produit ABIN6962281
  • Antigène Voir toutes NF-kB p65 (NFkBP65) Kits ELISA
    NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))
    Reactivité
    • 24
    • 20
    • 15
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    0.16 ng/mL - 10 ng/mL
    Seuil minimal de détection
    0.16 ng/mL
    Application
    ELISA
    Fonction
    The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.
    Type d'échantillon
    Cell Culture Supernatant, Plasma, Serum
    Analytical Method
    Quantitative
    Specificité
    This kit recognizes Human NFKB-p65 in samples. No Significant cross-reactivity or interference between Human NFKB-p65 and analogues was observed.
    Sensibilité
    0.1 ng/mL
    Ingrédients
    • Pre-coated, ready to use 96-well strip plate, flat buttom
    • Plate sealer for 96 wells
    • Reference Standard
    • Reference Standard & Sample Diluent
    • Biotinylated Detection Antibody (100 x concentrate)
    • HRP Conjugate (100 x concentrate)
    • Biotinylated Detection Antibody Diluent
    • HRP Conjugate Diluent
    • Substrate Reagent
    • Stop Solution
    • Wash Buffer (25 x concentrate)
    • Instruction manual
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  • Volume d'échantillon
    100 μL
    Durée du test
    3.5 h
    Plaque
    Pre-coated
    Protocole
    1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
    2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
    3. Aspirate and wash 3 times.
    4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
    5. Aspirate and wash 5 times.
    6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
    7. Add 50 µL Stop Solution. Read at 450 nm immediately.
    8. Calculation of results.
    Préparation des réactifs
    1. Bring all reagents to room temperature (18~25 °C) before use. Follow the Microplate reader manual for set-up and preheat it for 15 min before OD measurement.
    2. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer.Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved
    3. Standard working solution: Centrifuge the standard at 10,000xg for 1 min. Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 10 ng/mL. Then make serial dilutions as needed. The recommended dilution gradient is as follows: 10, 5, 2.5, 1.25, 0.63, 0.32, 0.16, 0 ng/mL. Dilution method: Take 7 EP tubes, add 500 μLof Reference Standard & Sample Diluent to each tube. Pipette 500 μLof the 10 ng/mL working solution to the first tube and mix up to produce a 5 ng/mL working solution. Pipette 500 μLof the solution from the former tube into the latter one according to these steps. The illustration below is for reference. Note: the last tube is regarded as a blank. Don't pipette solution into it from the former tube.
    4. Biotinylated Detection Antibody working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the stock tube before use, dilute the 100x Concentrated Biotinylated Detection Antibody to 1xworking solution with Biotinylated Detection Antibody Diluent.
    5. Concentrated HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Dilute the 100x Concentrated HRP Conjugate to 1x working solution with Concentrated HRP Conjugate Diluent.
    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): 3 samples with low, mid range and high level Human NFKB-p65 were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NFKB-p65 were tested on 3 different plates, 20 replicates in each plate.
    Both intra-CV and inter-CV are < 10 %.
    Restrictions
    For Research Use only
  • Stock
    4 °C,-20 °C
    Stockage commentaire
    1. For unopened kit: All reagents should be stored according to the labels on the vials, so they are stable up to 6 months after receipt of the kit. The Reference Standard, Biotinylated Detection Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
    2. For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.
    Date de péremption
    6 months
  • Yang, Liu, Liu, Ta: "Inhibition of TLR2/TLR4 alleviates the Neisseria gonorrhoeae infection damage in human endometrial epithelial cells via Nrf2 and NF-Kβsignaling." dans: Journal of reproductive immunology, Vol. 142, pp. 103192, (2020) (PubMed).

  • Antigène Voir toutes NF-kB p65 (NFkBP65) Kits ELISA
    NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))
    Autre désignation
    Nuclear factor NF-kappa-B p65 subunit (NFkBP65 Produits)
    Synonymes
    C-Rel Kit ELISA, NFKB3 Kit ELISA, p65 Kit ELISA, NFkB Kit ELISA, c-rel Kit ELISA, zgc:100833 Kit ELISA, Xrel2 Kit ELISA, Xrel3 Kit ELISA, rel Kit ELISA, rel-A Kit ELISA, rel2 Kit ELISA, rel3 Kit ELISA, v-rel Kit ELISA, xrel Kit ELISA, REL proto-oncogene, NF-kB subunit Kit ELISA, RELA proto-oncogene, NF-kB subunit Kit ELISA, v-rel reticuloendotheliosis viral oncogene homolog A (avian) Kit ELISA, v-rel avian reticuloendotheliosis viral oncogene homolog Kit ELISA, v-rel avian reticuloendotheliosis viral oncogene homolog L homeolog Kit ELISA, REL Kit ELISA, RELA Kit ELISA, Rela Kit ELISA, Rel Kit ELISA, rel Kit ELISA, rel.L Kit ELISA
    Sujet
    NFKB3, p65, RelA
    Pathways
    Signalisation NF-kappaB, Signalisation RTK, TCR Signaling, Signalisation TLR, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Hepatitis C, Toll-Like Receptors Cascades, S100 Proteins
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