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Clusterin ELISA Kit

CLU Reaktivität: Human Colorimetric Sandwich ELISA 15-20000 pg/mL Cell Culture Supernatant, Plasma, Serum
Produktnummer ABIN1979436
  • Target Alle Clusterin (CLU) ELISA Kits anzeigen
    Clusterin (CLU)
    Reaktivität
    • 7
    • 6
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    15-20000 pg/mL
    Untere Nachweisgrenze
    15 pg/mL
    Applikation
    ELISA
    Verwendungszweck
    Human Clusterin ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Proben
    Plasma, Cell Culture Supernatant, Serum
    Analytische Methode
    Quantitative
    Spezifität
    This ELISA antibody pair detect human Clusterin. Other species not determined.
    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 samples100,000 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 (Item E) should be used for dilution of serum/plasms/cell culture supernatants/urine. Suggested dilution for normal serum/plasma: 100,000 fold. The Human Clusterin ELISA Kit Protocol 3 For example, add 1 µL of serum/plasma into a tube with 199 µL 1x Assay Diluent to prepare a 200-fold diluted sample. Mix through and then pipette 1 µL of prepared 200-fold diluted sample into a tube with 499 µL 1x Assay Diluent to prepare a final 100,000 fold diluted sample. 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 (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 (Item E) into Item C vial to prepare a 80 ng/ml standard solution. Dissolve the powder thoroughly by a gentle mix. Add 150 µL Clusterin standard from the vial of Item C, into a tube with 450 µL 1x Assay Diluent to prepare a 20,000 pg/ml standard solution. Pipette 600myl 1x Assay Diluent into each tube. Use the 20,000 pg/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. 1x Assay Diluent serves as the zero standard (0 pg/ml). 200 µL 150 µL standard + 450 µL 200myl 200 µL 200 µL 200 µL 20,000 5,000 1,250 312.5 78.13 19.53 0 pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml The Human Clusterin ELISA Kit Protocol 4 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 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 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 1,000-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 12 µL of HRP-Streptavidin concentrate into a tube with 12 mL 1x Assay Diluent to prepare a 1,000-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 4) 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 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 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
  • Oztas, Ozler, Ersoy, Iskender, Sucak, Ergin, Uygur, Danisman: "Increased levels of serum clusterin is associated with intrauterine growth restriction and adverse pregnancy outcomes in preeclampsia." in: Journal of perinatal medicine, Vol. 44, Issue 3, pp. 269-75, (2016) (PubMed).

  • Target Alle Clusterin (CLU) ELISA Kits anzeigen
    Clusterin (CLU)
    Andere Bezeichnung
    Clusterin (CLU Produkte)
    Synonyme
    LOC100136120 ELISA Kit, clu ELISA Kit, MGC53055 ELISA Kit, zgc:73352 ELISA Kit, xclu ELISA Kit, MGC89163 ELISA Kit, CLU ELISA Kit, APO-J ELISA Kit, APOJ ELISA Kit, CLI ELISA Kit, CLU1 ELISA Kit, CLU2 ELISA Kit, KUB1 ELISA Kit, NA1/NA2 ELISA Kit, SGP-2 ELISA Kit, SGP2 ELISA Kit, SP-40 ELISA Kit, TRPM-2 ELISA Kit, TRPM2 ELISA Kit, DAG ELISA Kit, RATTRPM2B ELISA Kit, SP40 ELISA Kit, TRPM2B ELISA Kit, Trpm2 ELISA Kit, Trpmb ELISA Kit, AI893575 ELISA Kit, ApoJ ELISA Kit, Cli ELISA Kit, D14Ucla3 ELISA Kit, Sgp-2 ELISA Kit, Sgp2 ELISA Kit, Sugp-2 ELISA Kit, GP80 ELISA Kit, gpIII ELISA Kit, 40 ELISA Kit, clusterin-1 ELISA Kit, clusterin L homeolog ELISA Kit, clusterin ELISA Kit, Clusterin ELISA Kit, LOC100136120 ELISA Kit, clu.L ELISA Kit, clu ELISA Kit, CLU ELISA Kit, clus ELISA Kit, Clu ELISA Kit, LOC101113728 ELISA Kit
    Gen-ID
    1191
    UniProt
    P10909
    Pathways
    Apoptose, Negative Regulation of intrinsic apoptotic Signaling
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