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MUC1 Kit ELISA

MUC1 Reactivité: Humain Colorimetric Sandwich ELISA 4-1000 mU/mL Cell Culture Supernatant, Plasma, Serum
N° du produit ABIN625270
  • Antigène Voir toutes MUC1 Kits ELISA
    MUC1 (Mucin 1 (MUC1))
    Reactivité
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    4-1000 mU/mL
    Seuil minimal de détection
    4 mU/mL
    Application
    ELISA
    Fonction
    Human CA15-3 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Type d'échantillon
    Plasma, Cell Culture Supernatant, Serum
    Analytical Method
    Quantitative
    Specificité
    This ELISA kit shows no cross-reactivity with the following cytokines tested: human Angiogenin, BDNF, BLC, CNTF, ENA- 78, FGF-4, IL-1 alpha, IL-1 beta, 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-gamma, IGFBP-2, IGFBP-3, IGFBP-4, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIF, MIG, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, SDF-1 alpha, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    Sensibilité
    < 4 mU/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
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  • Indications d'application
    Recommended Dilution for serum and plasma samples30 - 200 fold
    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: 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 supernatants and urine. Suggested dilution for normal serum/plasma: 30-200 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 before use.
      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 supernates/urine) into Item C vial to prepare a 100 U/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 5 µL CA15-3 standard (100 U/mL) from the vial of Item C, into a tube with 495 µL Assay Diluent A or 1x Assay Diluent B to prepare a 1,000 mU/mL standard solution. Pipette 300 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the 1,000 mU/mL 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 mU/mL). 5 µL standard + 495 µL 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 1,000 400 160 64 25.6 10.24 4.10 0 mU/mL mU/mL mU/mL mU/mL mU/mL mU/mL mU/mL mU/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) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 200-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 50 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent B to prepare a 200-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    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.
      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. Assay Diluent A Human CA15-3 concentration (mU/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Human CA15-3 concentration (mU/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10
    Sensitivity: The minimum detectable dose of CA15-3 is typically less than 4 mU/mL.
    Recovery: Recovery was determined by spiking various levels human CA15-3 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 102.9 95-106 Plasma 107.6 100-115 Cell culture media 92.80 85-97
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93.03 98.78 96.45 Range ( %) 85-101 91-107 88-104 1:4 Average % of Expected 79.01 98.77 121.0 Range ( %) 71-87 78-138 110-132
    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 MUC1 Kits ELISA
    MUC1 (Mucin 1 (MUC1))
    Autre désignation
    CA15-3 (MUC1 Produits)
    Synonymes
    CA 15-3 Kit ELISA, CD227 Kit ELISA, EMA Kit ELISA, H23AG Kit ELISA, KL-6 Kit ELISA, MAM6 Kit ELISA, MCKD1 Kit ELISA, MUC-1 Kit ELISA, MUC-1/SEC Kit ELISA, MUC-1/X Kit ELISA, MUC1/ZD Kit ELISA, PEM Kit ELISA, PEMT Kit ELISA, PUM Kit ELISA, Muc-1 Kit ELISA, mucin 1, cell surface associated Kit ELISA, mucin 1, transmembrane Kit ELISA, MUC1 Kit ELISA, Muc1 Kit ELISA
    ID gène
    4582
    UniProt
    P15941
    Pathways
    Negative Regulation of intrinsic apoptotic Signaling
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