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Thrombopoietin ELISA Kit

THPO Reaktivität: Human Colorimetric Sandwich ELISA 90-6000 pg/mL Cell Culture Supernatant, Plasma, Serum
Produktnummer ABIN4884856
  • Target Alle Thrombopoietin (THPO) ELISA Kits anzeigen
    Thrombopoietin (THPO)
    Reaktivität
    • 5
    • 4
    • 4
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    90-6000 pg/mL
    Untere Nachweisgrenze
    90 pg/mL
    Applikation
    ELISA
    Verwendungszweck
    Human Thrombopoietin (TPO) 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 BDNF, BLC, ENA-78, FGF-4, IL-1alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, 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, PDGF, RANTES, SCF, TARC, TGF-beta1, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, VEGF.
    Sensitivität
    25 pg/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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  • 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) is used for dilution of serum/plasma samples and 1x Assay Diluent B (Item E) is used for dilution of culture supernatants and urine. 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. 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 40 µl TPO standard from the vial of Item C, into a tube with 626.7 µl Assay Diluent A or 1x Assay Diluent B to prepare a 6000 pg/ml stock standard solution. Pipette 400 µl Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Gently vortex to mix. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 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 80-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) before use. HRP-Streptavidin concentrate should be diluted 20,000-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 2 µl of HRP-Streptavidin concentrate into a tube with 198.0 µl 1x Assay Diluent B to prepare a 100-fold diluted HRP-Streptavidin solution (do not store the diluted solution for next day use). Mix through and then pipette 60 µl of prepared 100-fold diluted solution into a tube with 12 ml 1x Assay Diluent B to prepare a final 20,000 fold diluted HRP-Streptavidin solution.
    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 µl) 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 3. 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 3. 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.

    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • 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
  • Yilmaz, Basar, Altınbas, Ekiz, Aktas, Oztürk, Ginis, Coban, Ucar, Erarslan, Coskun, Yüksel, Tuna, Yüksel: "The utility of thrombopoietin in predicting liver fibrosis in chronic hepatitis B." in: International journal of clinical and experimental medicine, Vol. 7, Issue 5, pp. 1430-4, (2014) (PubMed).

    Sarkar, Schilffarth, Schams, Meyer, Berisha: "The expression of thrombopoietin and its receptor during different physiological stages in the bovine ovary." in: Reproduction in domestic animals = Zuchthygiene, Vol. 46, Issue 5, pp. 757-62, (2012) (PubMed).

  • Target Alle Thrombopoietin (THPO) ELISA Kits anzeigen
    Thrombopoietin (THPO)
    Andere Bezeichnung
    Thrombopoietin (THPO Produkte)
    Synonyme
    MGDF ELISA Kit, MKCSF ELISA Kit, ML ELISA Kit, MPLLG ELISA Kit, THCYT1 ELISA Kit, TPO ELISA Kit, Mgdf ELISA Kit, Ml ELISA Kit, Mpllg ELISA Kit, Tpo ELISA Kit, Tpo1 ELISA Kit, Tpo2 ELISA Kit, Tpo3 ELISA Kit, Tpo4 ELISA Kit, tpo ELISA Kit, LOC100231762 ELISA Kit, thrombopoietin ELISA Kit, THPO ELISA Kit, Thpo ELISA Kit, thpo ELISA Kit
    Hintergrund
    Human thrombopoietin (TPO), also known as megakaryocyte growth and development factor (MGDF) is 60-70 kDa. It is believed to play a key role in the development of megakaryocytes. TPO seems to stimulate an increase in megakaryocyte size and number. The Human TPO ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human TPO in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human TPO coated on a 96-well plate. Standards and samples are pipetted into the wells and TPO present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human TPO 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 TPO 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
    7066
    UniProt
    P40225
    Pathways
    JAK-STAT Signalweg, Hormone Activity
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