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Apelin Kit ELISA

APLN Reactivité: Souris Colorimetric Competition ELISA 98.77 pg/mL - 8000 pg/mL Plasma, Serum
N° du produit ABIN426254
  • Antigène Voir toutes Apelin (APLN) Kits ELISA
    Apelin (APLN)
    Reactivité
    • 4
    • 3
    • 2
    • 1
    • 1
    • 1
    Souris
    Méthode de détection
    Colorimetric
    Type de méthode
    Competition ELISA
    Gamme de detection
    98.77 pg/mL - 8000 pg/mL
    Seuil minimal de détection
    98.77 pg/mL
    Application
    ELISA
    Fonction
    The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of APLN in Serum,Plasma,Biological Fluids
    Type d'échantillon
    Plasma, Serum
    Analytical Method
    Quantitative
    Specificité

    This assay has high sensitivity and excellent specificity for detection of Apelin (APLN).
    No significant cross-reactivity or interference between Apelin (APLN) and analogues was observed.

    Réactivité croisée (Details)
    No significant cross-reactivity or interference between Apelin (APLN) and analogues was observed.
    Sensibilité
    35.3 pg/mL
    Ingrédients
    • Pre-coated, ready to use 96-well strip plate, flat buttom
    • Plate sealer for 96 wells
    • Reference Standard
    • Standard Diluent
    • Detection Reagent A
    • Detection Reagent B
    • Assay Diluent A
    • Assay Diluent B
    • Reagent Diluent (if Detection Reagent is lyophilized)
    • TMB Substrate
    • Stop Solution
    • Wash Buffer (30 x concentrate)
    • Instruction manual
    Matériel non inclus
    • Microplate reader with 450 nm filter.
    • Precision single or multi-channel pipettes and disposable tips.
    • Eppendorf Tubes for diluting samples.
    • Deionized or distilled water.
    • Absorbent paper for blotting the microtiter plate.
    • Container for Wash Solution
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  • Indications d'application
    • Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
    • The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
    • Kits from different batches may be a little different in detection range, sensitivity and color developing time.
    • Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
    • Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
    • There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
    • Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
    • Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
    • Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
    • Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.
    Commentaires

    Information on standard material:
    The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

    Information on reagents:
    The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

    Information on antibodies:
    The provided antibodies and their host vary in different kits.

    Volume d'échantillon
    50 μL
    Durée du test
    2 h
    Plaque
    Pre-coated
    Protocole
    1. Prepare all reagents, samples and standards,
    2. Add 50μL standard or sample to each well.
      Then add 50μL prepared Detection Reagent A immediately.
      Shake and mix. Incubate 1 hour at 37 °C,
    3. Aspirate and wash 3 times,
    4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
    5. Aspirate and wash 5 times,
    6. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
    7. Add 50μL Stop Solution. Read at 450 nm immediately.
    Préparation des réactifs
    • Bring all kit components and samples to room temperature (18-25°C) before use.
    • Standard - Reconstitute the Standard with 0.5mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 8,000pg/mL. Please prepare 5 tubes containing 0.6mL Standard Diluent and produce a triple dilution series. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 8,000pg/mL, 2,666.7pg/mL, 888.9pg/mL, 296.3pg/mL, 98.8pg/mL, and the last EP tubes with Standard Diluent is the blank as 0pg/mL.
    • Assay Diluent A and Assay Diluent B - Dilute 6mL of Assay Diluent A or B Concentrate(2×) with 6mL of deionized or distilled water to prepare 12 mL of Assay Diluent A or B. (In fact, more than 6mL Assay Diluent A and Assay Diluent B are contained in the bottles. Therefore, in every test, please precisely pipette required amount of Diluent and make double dilution in a new container. The prepared working dilution cannot be frozen.)
    • Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with working Assay Diluent A or B, respectively (1:100).
    • Wash Solution - Dilute 20mL of Wash Solution concentrate (30×) with 580mL of deionized or distilled water to prepare 600 mL of Wash Solution (1×).
    • TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.
    Note:
    • Making serial dilution in the wells directly is not permitted.
    • Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37°C directly.
    • Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
    • Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10µL for once pipetting.
    • The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
    • If crystals have formed in the Wash Solution concentrate (30×), warm to room temperature and mix gently until the crystals are completely dissolved.
    • Contaminated water or container for reagent preparation will influence the detection result.
    Prélèvement de l'échantillon
    Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

    Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

    Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.
    Calcul des résultats

    This assay employs the competitive inhibition enzyme immunoassay technique, so there is an inverse correlation between APLN concentration in the sample and the assay signal intensity. Average the duplicate readings for each standard, control, and samples. Create a standard curve on log-log or semi-log graph paper, with the log of APLN concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the standard points and it can be determined by regression analysis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
    In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the log of concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. The O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects). Typical standard curve below is provided for reference only.

    Précision du teste

    Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apelin (APLN) were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Apelin (APLN) were tested on 3 different plates, 8 replicates in each plate.
    CV(%) = SD/meanX100
    Intra-Assay: CV<10%
    Inter-Assay: CV<12%

    Restrictions
    For Research Use only
  • Précaution d'utilisation
    The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
    Conseil sur la manipulation
    The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
    To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
    Stock
    4 °C
    Stockage commentaire
    • For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
    • For opened kit: When the kit is opened, the remaining reagents still 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 reseal along entire edge of zip-seal.
      Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
    • For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.
    Date de péremption
    6 months
  • Li, Jiang, Yang, Wu, Sun, Sun: "CTRP3 modulates the expression and secretion of adipokines in 3T3-L1 adipocytes." dans: Endocrine journal, Vol. 61, Issue 12, pp. 1153-62, (2015) (PubMed).

    Stöhr, Kappel, Carnevale, Cavalera, Mavilio, Arisi, Fardella, Cifelli, Casagrande, Rizza, Cattaneo, Mauriello, Menghini, Lembo, Federici: "TIMP3 interplays with apelin to regulate cardiovascular metabolism in hypercholesterolemic mice." dans: Molecular metabolism, Vol. 4, Issue 10, pp. 741-52, (2015) (PubMed).

  • Antigène Voir toutes Apelin (APLN) Kits ELISA
    Apelin (APLN)
    Autre désignation
    APLN (APLN Produits)
    Synonymes
    APLN Kit ELISA, Xapelin Kit ELISA, xnpep2 Kit ELISA, preproapelin Kit ELISA, 6030430G11Rik Kit ELISA, Apel Kit ELISA, APEL Kit ELISA, XNPEP2 Kit ELISA, apelin Kit ELISA, apelin S homeolog Kit ELISA, apln Kit ELISA, APLN Kit ELISA, apln.S Kit ELISA, Apln Kit ELISA
    UniProt
    Q9R0R4
    Pathways
    Positive Regulation of Peptide Hormone Secretion, Hormone Activity, Feeding Behaviour
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