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EPOR ELISA Kit

EPOR Reaktivität: Human Colorimetric Sandwich ELISA 180-12.000 pg/mL Cell Culture Supernatant, Plasma, Serum
Produktnummer ABIN1979359
  • Target Alle EPOR ELISA Kits anzeigen
    EPOR (Erythropoietin Receptor (EPOR))
    Reaktivität
    • 3
    • 3
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    180-12.000 pg/mL
    Untere Nachweisgrenze
    180 pg/mL
    Applikation
    ELISA
    Verwendungszweck
    Human Erythropoietin R 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 the following cytokines tested: human Angiogenin, BDNF, BLC, CNTF, ENA-78, FGF-4, IL-1a, IL-1b, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, FGF-4, FGF-6, FGF-7, G-CSF, GDNF, GM-CSF, IFN-g, IGFBP-2, IGFBP-3, IGFBP-4, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIF, MIG, MIP-1a, MIP-1 b, MIP-1 delta, PARC, PDGF, RANTES, SCF, SDF-1 alpha, TARC, TGF-b, TIMP-1, TIMP-2, TNF-a, TNF-b, TPO, VEGF.
    Kreuzreaktivität (Details)
    This ELISA kit shows no cross-reactivity with the following cytokines tested: human Angiogenin, BDNF, BLC, CNTF, ENA- 78, FGF-4, IL-1alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL- 11, IL-12 p70, IL-12 p40, IL-13, IL-15, IL-309, IP-10, FGF-4, FGF-6, FGF- 7, G-CSF, GDNF, GM-CSF, IFN-gamma, IGFBP-2, IGF-BP-3, IGF-BP-4, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIF, MIG, MIP-1alpha, MIP-1 beta, MIP- 1delta, PARC, PDGF, RANTES, SCF,SDF-1alpha, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    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, 1x Assay Diluent D (Item K) should be used for dilution of serum/plasma /culture supernatants/urine. The Human EPO R ELISA Kit Protocol 3 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 D (Item K) and Assay Diluent B (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. Add 400 µL 1x Assay Diluent D (Item K, Assay Diluent D should be diluted 5-fold with deionized or distilled water before use) into Item C vial to prepare a 50 ng/ml standard solution. Dissolve the powder thoroughly by a gentle mix. Add 120 µL of the EPO R standard from the vial of Item C, into a tube with 380 µL 1x Assay Diluent D to prepare a 12,000 pg/ml standard solution. Pipette 250 µL 1x Assay Diluent D into each tube. Use 12,000 pg/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. 1x Assay Diluent D serves as the zero standard (0 pg/ml). 200 µL 200 µL 200 µL 200 µL 200 µL 120 µL standard + 380 µL 200myl 12,000 6,000 3,000 1,500 750 375 187.5 0 pg/ml 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 The Human EPO R ELISA Kit Protocol 4 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 (Item E) 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) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 300-fold with 1x Assay Diluent B (Item E). 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 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 overnight at 4 °C with gentle shaking. 3. Discard 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.

    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
  • Target Alle EPOR ELISA Kits anzeigen
    EPOR (Erythropoietin Receptor (EPOR))
    Andere Bezeichnung
    EPO R (EPOR Produkte)
    Synonyme
    EPOR ELISA Kit, epor ELISA Kit, EPO-R ELISA Kit, erythropoietin receptor ELISA Kit, erythropoietin receptor L homeolog ELISA Kit, EPOR ELISA Kit, epor ELISA Kit, Epor ELISA Kit, epor.L ELISA Kit
    Gen-ID
    2057
    UniProt
    P19235
    Pathways
    JAK-STAT Signalweg
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