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HGF Kit ELISA

HGF Reactivité: Souris Colorimetric Sandwich ELISA Cell Lysate, Tissue Lysate
N° du produit ABIN625127
  • Antigène Voir toutes HGF Kits ELISA
    HGF (Hepatocyte Growth Factor (Hepapoietin A, Scatter Factor) (HGF))
    Reactivité
    • 8
    • 6
    • 5
    • 4
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Souris
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Application
    ELISA
    Fonction
    Mouse HGF ELISA Kit for cell and tissue lysate samples.
    Type d'échantillon
    Cell Lysate, Tissue Lysate
    Analytical Method
    Quantitative
    Specificité
    The antibody pair provided in this kit recognizes mouse HGF.
    Sensibilité
    400 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 (Item D) and Assay Diluent (Item E) should be diluted 5-fold with deionized or distilled water before use.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL 1x Sample Diluent Buffer into Item C vial to prepare a 60 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Pipette 240 µL Sample Diluent Buffer into each tube. Use the 60 ng/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Sample Diluent Buffer serves as the zero standard (0 ng/mL). Item C vial + 400 µL 160 µL 160 µL 160 µL 160 µL 160myl 60 24 9.6 3.84 1.536 0.614 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL
      5. If 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 80-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 500-fold with 1x Assay Diuent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 10 m µL 1x Assay Diluent to prepare a 500-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
      8. Cell lysate buffer should be diluted 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 and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking. We recommend using 50-500 myg/mL of total protein for lysate sample. The amount of sample used depends on the abundance of target protein. More of the sample can be used if signals are too weak. If signals are too strong, the sample can be diluted further.
      3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
      4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
      5. Discard the solution. Repeat the wash as in step
      6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
      7. Discard the solution. Repeat the wash as in step
      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 with gentle shaking.
      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.
    Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Sample Assay Diluent Mouse HGF concentration (ng/mL) 0.1 1 10 100 O D =4 50 n m 0.001 0.01 0.1 1 10
    Sensitivity: The minimum detectable dose of HGF is typically less than 400 pg/mL.
    Recovery: Recovery was determined by spiking various levels of mouse HGF into mouse tissue lysate and cell lysate. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Tissue lysate 120.2 110-130 Cell lysate 114.4 100-134
    Linearity: Sample Type Tissue Cell Lysate lysate 1:2 Average % of 118.9 139.4 Expected Range ( %) 109-125 130-143 1:4 Average % of 132.4 129.2 Expected Range ( %) 120-144 120-137
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Précision du teste
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Restrictions
    For Research Use only
  • Conseil sur la manipulation
    Avoid repeated freeze-thaw cycles.
    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 HGF Kits ELISA
    HGF (Hepatocyte Growth Factor (Hepapoietin A, Scatter Factor) (HGF))
    Autre désignation
    HGF (HGF Produits)
    Synonymes
    DFNB39 Kit ELISA, F-TCF Kit ELISA, HGFB Kit ELISA, HPTA Kit ELISA, SF Kit ELISA, HGF Kit ELISA, C230052L06Rik Kit ELISA, HGF/SF Kit ELISA, NK1 Kit ELISA, NK2 Kit ELISA, SF/HGF Kit ELISA, hepatocyte growth factor Kit ELISA, HGF Kit ELISA, Hgf Kit ELISA
    Sujet
    The Mouse HGF (Hepatocyte Growth Factor) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse HGF cell lysate and tissue lysate. This assay employs an antibody specific for mouse HGF coated on a 96-well plate. Standards and samples are pipetted into the wells and HGF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse HGF 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 HGF 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
    15234
    UniProt
    Q08048
    Pathways
    Signalisation RTK, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Synaptic Membrane, Signaling of Hepatocyte Growth Factor Receptor
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