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FGF9 ELISA Kit

FGF-9 Reaktivität: Human Colorimetric Sandwich ELISA 200-35000 pg/mL Cell Culture Supernatant, Plasma, Serum
Produktnummer ABIN624979
  • Target Alle FGF9 (FGF-9) ELISA Kits anzeigen
    FGF9 (FGF-9) (Fibroblast Growth Factor 9 (FGF-9))
    Reaktivität
    • 3
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    200-35000 pg/mL
    Untere Nachweisgrenze
    200 pg/mL
    Applikation
    ELISA
    Verwendungszweck
    Human FGF-9 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Proben
    Plasma, Cell Culture Supernatant, Serum
    Analytische Methode
    Quantitative
    Spezifität
    This ELISA kit shows no cross-reactivity with any of the following cytokines tested: human Angiogenin, BDNF, BLC, ENA-78, FGF- 4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    Sensitivität
    < 200pg/mL
    Produktmerkmale
    • 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
    Bestandteile
    • 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
    Benötigtes Material
    • 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
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    Discover our top product FGF-9 ELISA Kit
  • Applikationshinweise
    Recommended Dilution for serum and plasma samples2 fold
    Probenmenge
    100 μL
    Plattentyp
    Pre-coated
    Protokoll
    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.
    Aufbereitung der Reagenzien
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
      2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernantants and urine. Suggested dilution for normal serum/plasma: 2 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
      3. Assay Diluent B should be diluted 5-fold with deionized or distilled water.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 100 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 200 µL FGF-9 standard from the vial of Item C, into a tube with 371.4 µL Assay Diluent A or 1x Assay Diluent B to prepare a 35,000 pg/mL stock standard solution. Pipette 300 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 200myl 200 µL 200 µL 200 µL 200myl standard + 371myl 30,000 35,000 14,000 5,600 2,240 896.0 358.4 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/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 Diluent B 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 Diluent B and used in step 4 of Part VI Assay Procedure.
      7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 300-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 40 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a final 300 fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    Testdurchführung
    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.
      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.
    Ergebnisberechnung

    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. Assay Diluent A Human FGF-9 concentrations (ng/mL) O D =4 50 n m 0.1 1 10 0 0.1 1 10 100 Assay Diluent B Human FGF-9 concentrations (ng/mL) O D =4 50 n m 0.01 0.1 1 10 0 0.1 1 10 100
    Sensitivity: The minimum detectable dose of FGF-9 is typically less than 200pg/mL
    Recovery: Recovery was determined by spiking various levels of human FGF-9 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 97.52 85-105 Plasma 94.38 83-102 Cell culture media 100.27 88-107
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 95 94 97 Range ( %) 86-105 82-102 84-103 1:4 Average % of Expected 97 96 101 Range ( %) 87-106 88-105 89-108
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Testpräzision
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Handhabung
    Avoid repeated freeze-thaw cycles.
    Lagerung
    -20 °C
    Informationen zur Lagerung
    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.
    Haltbarkeit
    6 months
  • Suzuki, Yasuda, Funaishi, Arai, Ishioka, Ohgino, Tani, Hamamoto, Ohashi, Naoki, Betsuyaku, Soejima: "Multiple roles of extracellular fibroblast growth factors in lung cancer cells." in: International journal of oncology, Vol. 46, Issue 1, pp. 423-9, (2014) (PubMed).

    Naruo, Seko, Kuroshima, Matsutani, Sasada, Kondo, Kurokawa: "Novel secretory heparin-binding factors from human glioma cells (glia-activating factors) involved in glial cell growth. Purification and biological properties." in: The Journal of biological chemistry, Vol. 268, Issue 4, pp. 2857-64, (1993) (PubMed).

  • Target Alle FGF9 (FGF-9) ELISA Kits anzeigen
    FGF9 (FGF-9) (Fibroblast Growth Factor 9 (FGF-9))
    Andere Bezeichnung
    FGF9 (FGF-9 Produkte)
    Synonyme
    Eks ELISA Kit, FGF-9 ELISA Kit, GAF ELISA Kit, HBFG-9 ELISA Kit, HBGF-9 ELISA Kit, SYNS3 ELISA Kit, XFGF-9 ELISA Kit, fgf9-A ELISA Kit, fibroblast growth factor 9 ELISA Kit, fibroblast growth factor 9 S homeolog ELISA Kit, Fgf9 ELISA Kit, FGF9 ELISA Kit, fgf9.S ELISA Kit
    Hintergrund
    FGF-9 (Fibroblast growth factor-9) is a member of the fibroblast growth factor (FGF) family of proteins. Human FGF-9 is a protein of 208 amino acids with sequence similarity of approximately 30 percent to other members of the family of FGF. FGF-9 is found in the conditioned medium of a human glioma cell line and acts on cells of the central nervous system. It is a potent mitogen for glial cells. The Human FGF-9 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human FGF-9 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human FGF-9 coated on a 96-well plate. Standards and samples are pipetted into the wells and FGF-9 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human FGF-9 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-9 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%.
    Gen-ID
    2254
    UniProt
    P31371
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