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FGF4 Kit ELISA

FGF4 Reactivité: Humain Colorimetric Sandwich ELISA Cell Culture Supernatant, Plasma, Serum
N° du produit ABIN624976
  • Antigène Voir toutes FGF4 Kits ELISA
    FGF4 (Fibroblast Growth Factor 4 (FGF4))
    Reactivité
    • 2
    • 2
    • 2
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Application
    ELISA
    Fonction
    Human FGF-4 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Type d'échantillon
    Plasma, Cell Culture Supernatant, Serum
    Analytical Method
    Quantitative
    Specificité
    The antibody pair provided in this kit recognizes human FGF-4.
    Sensibilité
    50 pg/mL
    Attributs du produit
    • Strip plates and additional reagents allow for use in multiple experiments
    • Quantitative protein detection
    • Establishes normal range
    • The best products for confirmation of antibody array data
    Ingrédients
    • Pre-Coated 96-well Strip Microplate
    • Wash Buffer
    • Stop Solution
    • Assay Diluent(s)
    • Lyophilized Standard
    • Biotinylated Detection Antibody
    • Streptavidin-Conjugated HRP
    • TMB One-Step Substrate
    Matériel non inclus
    • Distilled or deionized water
    • Precision pipettes to deliver 2 μL to 1 μL volumes
    • Adjustable 1-25 μL pipettes for reagent preparation
    • 100 μL and 1 liter graduated cylinders
    • Tubes to prepare standard and sample dilutions
    • Absorbent paper
    • Microplate reader capable of measuring absorbance at 450nm
    • Log-log graph paper or computer and software for ELISA data analysis
    • Cell lysate buffer
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  • Volume d'échantillon
    100 μL
    Plaque
    Pre-coated
    Protocole
    1. Prepare all reagents, samples and standards as instructed in the manual.
    2. Add 100 μL of standard or sample to each well.
    3. Incubate 2.5 h at RT or O/N at 4 °C.
    4. Add 100 μL of prepared biotin antibody to each well.
    5. Incubate 1 h at RT.
    6. Add 100 μL of prepared Streptavidin solution to each well.
    7. Incubate 45 min at RT.
    8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 μL of Stop Solution to each well.
    11. Read at 450 nm immediately.
    Préparation des réactifs
    1. Bring all reagents and samples to room temperature (18 - 25°C) before use. 2. Sample dilution: Tissue lysate and cell lysate sample should be diluted at least 5-fold with 1x Sample Diluent Buffer. 3. Sample Diluent Buffer/Assay Diluent should be diluted 5-fold with deionized or distilled water. 4. paration of standard: Briefly spin the vial of Item C. Add 400 µl 1x Sample Diluent Buffer(Item D) into item C vial to prepare a 50 ng/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 180 µl FGF-4 standard from the vial of Item C, into a tube with 320 µl 1x Sample Diluent Buffer to prepare a 18,000 pg/ml stock standard Solution. Pipette 400 µl 1x Sample Diluent Buffer into each tube. Use the stock standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. 1x Assay Diluent 1x Sample Diluent Buffer serves as the zero standard (0 pg/ml). 5. the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer. 6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µl of 1x Assay Diuent into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4°C for 5 days). The detection antibody concentrate should be diluted 65-fold with 1x Assay Diuent and used in step 4 of Part VI Assay Procedure. 7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) before use. HRP-Streptavidin concentrate should be diluted 6,000-fold with 1x Assay Diuent. 8. Cell lysate buffer is diluted to 2-fold with deionized or distilled water (for cell lysate and tissue lysate).
    Procédure de l'essai
    1. Bring all reagents and samples to room temperature (18 - 25°C) before use. It is recommended that all standards and samples be run at least in duplicate. 2. Add 100 µl of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well with plate holder and incubate for 2.5 hours at room temperature or over night at 4°C. 3. Discard the solution and wash 4 times with 1x Wash Solution (200 µl each). 4. Add 100 µl of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature. 5. Discard the solution and wash 4 times with 1x Wash Solution (200 µl each). 6. Add 100 µl of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature. 7. Discard the solution and wash 5 times with 1x Wash Solution (200 µl each). 8. Add 100 µl of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark. 9. Add 50 µl of Stop Solution (Item I) to each well. Read at 450 nm immediately.
    Calcul des résultats

    Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.

    Restrictions
    For Research Use only
  • Stock
    -20 °C
    Stockage commentaire
    The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
    Date de péremption
    6 months
  • Antigène Voir toutes FGF4 Kits ELISA
    FGF4 (Fibroblast Growth Factor 4 (FGF4))
    Autre désignation
    FGF-4 (FGF4 Produits)
    Synonymes
    HBGF-4 Kit ELISA, HST Kit ELISA, HST-1 Kit ELISA, HSTF1 Kit ELISA, K-FGF Kit ELISA, KFGF Kit ELISA, Fgf-4 Kit ELISA, Fgfk Kit ELISA, Hst1 Kit ELISA, Hstf-1 Kit ELISA, KS3 Kit ELISA, hst Kit ELISA, hst-1 Kit ELISA, kFGF Kit ELISA, Hst Kit ELISA, fibroblast growth factor 4 Kit ELISA, FGF4 Kit ELISA, Fgf4 Kit ELISA
    Sujet
    FGF (Fibroblast growth factors) constitutes a family of related 16-18 kDa proteins controlling normal growth and differentiation of mesenchymal, epithelial, and neuroectodermal cell types. FGF-4 has potent transforming potential, apparently through an autocrine mechanism of action, and it is a potent angiogenic factor. It has also been extensively studied for its effects in embryogenesis. The Human FGF-4 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human FGF-4 cell lysate and tissue lysate. This assay employs an antibody specific for human FGF-4 coated on a 96-well plate. Standards and samples are pipetted into the wells and FGF-4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human FGF-4 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of FGF-4 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
    ID gène
    2249
    UniProt
    P08620
    Pathways
    Signalisation RTK, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Stem Cell Maintenance
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