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CX3CL1 ELISA Kit

CX3CL1 Reaktivität: Ratte Colorimetric Sandwich ELISA Cell Lysate, Tissue Lysate
Produktnummer ABIN625193
  • Target Alle CX3CL1 ELISA Kits anzeigen
    CX3CL1 (Chemokine (C-X3-C Motif) Ligand 1 (CX3CL1))
    Reaktivität
    • 6
    • 5
    • 4
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Ratte
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Applikation
    ELISA
    Verwendungszweck
    Rat Fractalkine (CX3CL1) ELISA Kit for cell and tissue lysate samples.
    Proben
    Cell Lysate, Tissue Lysate
    Analytische Methode
    Quantitative
    Spezifität
    The antibody pair provided in this kit recognizes rat Fractalkine.
    Kreuzreaktivität (Details)
    This ELISA kit shows no cross-reactivity with any of the cytokines tested (e.g., rat CINC-2, CINC-3, CNTF, IL-1alpha, IL-1beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3alpha, beta-NGF, TIMP-1, TNF-alpha, VEGF)
    Sensitivität
    5 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
    • Cell lysate buffer
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  • 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: Tissue lysate and cell lysate sample should be diluted at least 5-fold with 1x Sample Diluent Buffer.
      3. Sample Diluent Buffer (Item D) and 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 Sample Diluent Buffer (Item D) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL Fractalkine standard from the vial of Item C, into a tube with 960 µL Sample Diluent Buffer to prepare a 2,000 pg/mL stock standard solution. Pipette 400 µL 1x Sample Diluent Buffer into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Sample Diluent Buffer serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200myl 200 µL 200 µL 40 µL standard + 960 µL 2000 666.7 222.2 74.07 24.69 8.23 2.74 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 Diuent 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 Diuent 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 240-fold with 1x Assay Diuent. 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 12 ml 1x Assay Diluent to prepare a 240-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.
      8. Cell lysate buffer should be diluted 2-fold with deionized or distilled water (for cell lysate and tissue lysate).
    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. We recommend using 50-500 myg/mL of total protein for lysate sample. The amount of sample used depends on the abundance of target protein. More of the sample can be used if signals are too weak. If signals are too strong, the sample can be diluted further.
      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 Rat Fractalkine concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 (n m ) 0.01 0.1 1 10
    Sensitivity: The minimum detectable dose of Fractalkine is typically less than 5 pg/mL.
    Recovery: Recovery was determined by spiking various levels of rat Fractalkine into cell lysis buffer. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Tissue lysate 92.87 83-103 Cell lysate 93.54 84-104
    Linearity: Sample Type Tissue Cell Lysate lysate 1:2 Average % of 89 91 Expected Range ( %) 81-101 82-102 1:4 Average % of 92 96 Expected Range ( %) 84-103 83-103
    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
  • Target Alle CX3CL1 ELISA Kits anzeigen
    CX3CL1 (Chemokine (C-X3-C Motif) Ligand 1 (CX3CL1))
    Andere Bezeichnung
    Fractalkine / CX3CL1 (CX3CL1 Produkte)
    Synonyme
    ABCD3 ELISA Kit, CX3CL1 ELISA Kit, DKFZp459K117 ELISA Kit, Cx3c ELISA Kit, Scyd1 ELISA Kit, ABCD-3 ELISA Kit, C3Xkine ELISA Kit, CXC3 ELISA Kit, CXC3C ELISA Kit, NTN ELISA Kit, NTT ELISA Kit, SCYD1 ELISA Kit, fractalkine ELISA Kit, neurotactin ELISA Kit, AB030188 ELISA Kit, AI848747 ELISA Kit, CX3C ELISA Kit, Cxc3 ELISA Kit, D8Bwg0439e ELISA Kit, ATP binding cassette subfamily D member 3 ELISA Kit, C-X3-C motif chemokine ligand 1 ELISA Kit, chemokine (C-X3-C motif) ligand 1 ELISA Kit, ABCD3 ELISA Kit, CX3CL1 ELISA Kit, Cx3cl1 ELISA Kit, Abcd3 ELISA Kit
    Hintergrund
    Chemokine (C-X3-C motif) ligand 1 (Cx3cl1 protein) (Fractalkine)
    UniProt
    Q6IRF7
    Pathways
    Synaptic Membrane
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