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CXCL5 ELISA Kit

CXCL5 Reaktivität: Human Colorimetric Sandwich ELISA 10-18000 pg/mL Cell Culture Supernatant, Plasma, Serum
Produktnummer ABIN624966
  • Target Alle CXCL5 ELISA Kits anzeigen
    CXCL5 (Chemokine (C-X-C Motif) Ligand 5 (CXCL5))
    Reaktivität
    • 7
    • 4
    • 4
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    10-18000 pg/mL
    Untere Nachweisgrenze
    10 pg/mL
    Applikation
    ELISA
    Verwendungszweck
    Human ENA-78 (CXCL5) 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 cytokines tested: Human Angiogenin, BDNF, BLC, 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-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-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, MMP-1, -2, -7, -8, -9, -10, -12, -13, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    Sensitivität
    < 10 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 - 10 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 should be used for dilution of serum/plasma samples. 1x Assay Diluent B should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 2-10 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 120 µL ENA-78 standard from the vial of Item C, into a tube with 546.7 µL Assay Diluent A or 1x Assay Diluent B to prepare a 18000 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 . 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 120 µL standard +546.7 µL 200myl 200 µL 200 µL 200 µL 200 µL 18000 6000 2000 666.7 222.2 74.07 24.69 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 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 500-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent B to prepare a final 500 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 ENA-78 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 10 100 1,000 10,000 100,000 Assay Diluent B Human ENA-78 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 10 100 1,000 10,000 100,000
    Sensitivity: The minimum detectable dose of ENA-78 is typically less than 10 pg/mL.
    Recovery: Recovery was determined by spiking various levels of human ENA-78 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 93.39 82-102 Plasma 92.15 81-101 Cell culture media 102.15 86-106
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93 94 94 Range ( %) 82-102 83-102 83-102 1:4 Average % of Expected 94 93 96 Range ( %) 83-104 82-102 84-103
    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
  • Spaks, Jaunalksne, Spaka, Chudasama, Pirtnieks, Krievins: "Diagnostic Value of Circulating CXC Chemokines in Non-small Cell Lung Cancer." in: Anticancer research, Vol. 35, Issue 12, pp. 6979-83, (2016) (PubMed).

    Wang, Su, Yang, Qiao, Fang, Yu, Yang, Wang, Yin, Chen, Hong: "The influence of myeloid-derived suppressor cells on angiogenesis and tumor growth after cancer surgery." in: International journal of cancer, Vol. 138, Issue 11, pp. 2688-99, (2016) (PubMed).

    Weber: "[Metal ceramic boundary surfaces in the Ni-Cr-ceramic system from a recent viewpoint]." in: Deutsche zahnärztliche Zeitschrift, Vol. 37, Issue 11, pp. 896-9, (1983) (PubMed).

  • Target Alle CXCL5 ELISA Kits anzeigen
    CXCL5 (Chemokine (C-X-C Motif) Ligand 5 (CXCL5))
    Andere Bezeichnung
    ENA78 (CXCL5 Produkte)
    Synonyme
    ENA-78 ELISA Kit, SCYB5 ELISA Kit, Cxcl6 ELISA Kit, CXCL5 ELISA Kit, GCP-2 ELISA Kit, GCP2 ELISA Kit, SCYB6 ELISA Kit, AMCF-II ELISA Kit, LIX ELISA Kit, Scyb5 ELISA Kit, Scyb6 ELISA Kit, C-X-C motif chemokine ligand 5 ELISA Kit, C-X-C motif chemokine ligand 6 ELISA Kit, chemokine (C-X-C motif) ligand 5 ELISA Kit, C-X-C motif chemokine 5 ELISA Kit, CXCL5 ELISA Kit, Cxcl6 ELISA Kit, LOC701946 ELISA Kit, LOC703222 ELISA Kit, Cxcl5 ELISA Kit
    Hintergrund
    The Human ENA-78 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human ENA-78 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human ENA-78 coated on a 96-well plate. Standards and samples are pipetted into the wells and ENA-78 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human ENA-78 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 ENA-78 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
    6374
    UniProt
    P42830
    Pathways
    Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity
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