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IL-8 Kit ELISA

IL8 Reactivité: Humain Colorimetric Sandwich ELISA 16-1000 pg/mL Cell Lysate, Plasma, Serum
N° du produit ABIN924784
  • Antigène Voir toutes IL-8 (IL8) Kits ELISA
    IL-8 (IL8) (Interleukin 8 (IL8))
    Reactivité
    • 14
    • 5
    • 4
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    16-1000 pg/mL
    Seuil minimal de détection
    16 pg/mL
    Application
    ELISA
    Fonction
    The OmniKine? Human IL-8 ELISA Kit contains the components necessary for quantitative determination of natural or recombinant Human IL-8 concentrations within any experimental sample including cell lysates, serum and plasma. This particular immunoassay utilizes the quantitative technique of a "Sandwich" Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a "sandwich" format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on Human IL-8 while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non- specific binding between proteins to other proteins or to the solid phase. After incubation and "sandwiching" of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a colorimetric reaction to ensue upon substrate addition. When the substrate TMB (3, 3', 5, 5'-Tetramethylbenzidine) is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution (2 N Sulfuric Acid) where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.
    Marque
    OmniKine™
    Type d'échantillon
    Cell Lysate, Serum, Plasma
    Analytical Method
    Quantitative
    Specificité
    The Human IL-8 ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human IL-8 proteins.
    Réactivité croisée (Details)
    The Human IL-8 ELISA is capable of recognizing both recombinant and naturally produced Human IL-8 proteins. The antigens listed below were tested at 50 ng/mL and exhibited 100% cross reactivity. Human: IL-8 (77 aa) The antigens listed below were tested at 50 ng/mL and did not exhibit significant cross reactivity or interference. Human: BCA-1, BRAK, CXCL-16, ENA-78, Fractalkine, GCP-2, GRO, GRO-β, GRO-γ, I-TAC, IP-10, Lymphotactin, MCP-1, MIG, NAP-2, PF-4, SDF-1α, SDF-1β
    Attributs du produit
    The Human IL-8 ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human IL-8 proteins within the range of 16-1000 pg/mL.
    Ingrédients
    • Microstrips Coated w / Capture Antibody: 12 x 8-Well Microstrips
    • Protein Standard: Lyophilized (100 ng), Red container
    • Biotinylated Detection Antibody: Lyophilized, Yellow container
    • 400x Streptavidin-HRP: 30 μL, Blue container
    • Wash Buffer (10x): 50 mL, Clear containter
    • Assay Diluent: 50 mL, Clear container
    • Ready-to-Use Substrate: 12 mL, Brown container
    • Stop Solution: 12 mL, Clear container
    • Adhesive Plate Sealers: 4 Sheets
    • Technical Manual 1 Manual
    Matériel non inclus
    The following materials and/or equipment are NOT provided in this kit but are necessary to successfully conduct the experiment:
    Microplate reader able to measure absorbance at 450 nm (with correction wavelength set to 540 nm or 570 nm)
    Micropipettes with capability of measuring volumes ranging from 1 μl to 1 mL
    Deionized or sterile water
    Squirt bottle, manifold dispenser, multichannel pipette reservoir or automated microplate washer
    Graph paper or computer software capable of generating or displaying logarithmic functions
    Absorbent paper or vacuum aspirator
    Test tubes or microfuge tubes capable of storing ≥1 mL
    Bench
    top centrifuge (optional)
    Bench
    top vortex (optional)
    Orbital shaker (optional)
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  • Plaque
    Pre-coated
    Protocole
    This particular immunoassay utilizes the quantitative technique of a Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a sandwich format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on the Human IL-8 cytokine while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non-specific binding between proteins to other proteins or to the solid phase. After incubation and sandwiching of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a colorimetric reaction to ensue upon substrate addition. When the substrate TMB (3, 3’, 5, 5’- Tetramethylbenzidine) is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution (2 N Sulfuric Acid) where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.
    Préparation de l'échantillon

    If samples are to be used within 24 hours, aliquot and store at 4 °C. If samples are to be used over a long period of time, aliquot and store between -20 °C and -80 °C, depending on the duration of storage.
    Note: Samples containing a visible precipitate or pellet must be clarified prior to use in the assay.
    Caution: Avoid repeated freeze/thaw cycles to prevent loss of biological activity of proteins in experimental samples.

    • Cell Lysate and Supernatants:
      Remove large cell components via centrifugation and perform the assay. Cell lysates and supernatants require a dilution using Assay Diluent. A serial dilution may be performed to determine a suitable dilution factor for the sample. For future use of the sample, follow the sample storage guidelines stated above.
    • Serum:
      Allow samples to clot in a serum separator tube (SST) for 30 minutes. After sufficient clotting, centrifuge at 1000 x g for 15 minutes and remove serum from SST in preparation for the assay. Serum samples require at least a 1:50 dilution using Assay Diluent. For future use of the sample, follow the storage guidelines above.
    • Plasma:
      Use heparin, citrate or EDTA as an anticoagulant to gather plasma from original biological sample. After collection of the plasma, centrifuge for 15 minutes at 1000 x g. This step must be performed within 30 minutes of plasma collection. Plasma samples require at least a 1:50 dilution using Assay Diluent. Afterwards, perform the assay or for future use of the sample, follow the storage guidelines stated above.

    Procédure de l'essai

    Note: If possible, all incubation steps should be performed on an orbital shaker to equilibrate solutions when added to the microplate wells. Also, all provided solutions should be at ambient temperature prior to use.
    Note: Avoid adding solutions into wells at an angle, always keep pipette tip perpendicular to plate bottom.

    Reconstitution of Provided Materials:

      1. Reconstitute the Biotin-Conjugated Detection Antibody in 67 µL of ddH₂O for a concentration of 180 µg/ml.
      2. Reconstitute the Protein Standard in 100 µL of ddH₂O for a concentration of 340 ng/ml.
      3. Dilute the 50 mL of 10x Wash Buffer in 450 mL of ddH2O for 500 mL of 1x Wash Buffer.
    Addition of Known Standard and Unknown Sample to Immunoassay:
      The OmniKine™ Human CD163 ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human CD163 proteins

    Calcul des résultats

    Generation of Standard Curve and Interpretation of Data
    1. Average the duplicate or triplicate readings for each standard, control and sample and subtract the average zero standard optical density.
    2. Generate a standard curve by using Microsoft Excel or other computer software capable of establishing a 4- Parameter Logistic (4-PL) curve fit. If using Excel or an alternative graphing tool, plot the average optical density values in absorbance units (y-axis) against the known standard concentrations in pg/ml (x-axis). Note: Only use the values in which a noticeable gradient can be established. Afterwards, generate a best fit curve or trend-line through the plotted points via regression analysis.

    Restrictions
    For Research Use only
  • Précaution d'utilisation
    Reagents provided in this kit may be harmful if ingested, inhaled or absorbed through the skin. Please carefully review the MSDS for each reagent before conducting the experiment.
    Stop Solution contains 2 N Sulfuric Acid (H2SO4) and is an extremely corrosive agent. Please wear proper eye, hand and face protection when handling this material. When the experiment is finished, be sure to rinse the plate with copious amounts of running water to dilute the Stop Solution prior to disposing the plate.
    Conseil sur la manipulation
    This ELISA kit is intended for research purposes only, NOT diagnostic or clinical procedures of any kind.
    Materials included in this kit should NOT be used past the expiration date on the kit label.
    Reagents or substrates included in this kit should NOT be mixed or substituted with reagents or substrates from any other kits.
    Variations in pipetting technique, washing technique, operator laboratory technique, kit age, incubation time or temperature may cause differences in binding affinity of the materials provided.
    The assay is designed to eliminate interference and background by other cellular macromolecules or factors present within any biological samples. However, the possibility of background noise cannot be fully excluded until all factors have been tested using the assay kit.

    Reagents provided in this kit may be harmful if ingested, inhaled or absorbed through the skin. Please carefully review the MSDS for each reagent before conducting the experiment.
    Stop Solution contains 2 N Sulfuric Acid (H2SO4) and is an extremely corrosive agent. Please wear proper eye, hand and face protection when handling this material. When the experiment is finished, be sure to rinse the plate with copious amounts of running water to dilute the Stop Solution prior to disposing the plate.
    Stock
    4 °C
    Stockage commentaire
    Note: If used frequently, reagents may be stored at 4 °C.
    • Unopened Kits: Store at 4 °C for 6 months.
    • Microstrips Coated w/ Capture Antibody, 400x Streptavidin-HRP Wash Buffer (10x), Assay Diluent Ready-to-Use Substrate, Stop Solution: 6 Months at 4 °C
    • Protein Standard, Biotinylated Detection Antibody: Lyophilized: 6 Months (if Reconstituted: 1 Month) at 4 °C
  • Antigène Voir toutes IL-8 (IL8) Kits ELISA
    IL-8 (IL8) (Interleukin 8 (IL8))
    Autre désignation
    IL-8 (IL8 Produits)
    Synonymes
    CXCL8 Kit ELISA, GCP-1 Kit ELISA, GCP1 Kit ELISA, LECT Kit ELISA, LUCT Kit ELISA, LYNAP Kit ELISA, MDNCF Kit ELISA, MONAP Kit ELISA, NAF Kit ELISA, NAP-1 Kit ELISA, NAP1 Kit ELISA, IL-8 Kit ELISA, AMCF-I Kit ELISA, il8 Kit ELISA, IL8 Kit ELISA, si:dkey-151b16.2 Kit ELISA, cxcli2 Kit ELISA, cxcl8 Kit ELISA, gcp-1 Kit ELISA, il-8 Kit ELISA, lynap Kit ELISA, mdncf Kit ELISA, monap Kit ELISA, nap-1 Kit ELISA, C-X-C motif chemokine ligand 8 Kit ELISA, interleukin 8 Kit ELISA, chemokine (C-X-C motif) ligand 8a Kit ELISA, chemokine (C-X-C motif) ligand 1-like Kit ELISA, chemokine (C-X-C motif) ligand 8 L homeolog Kit ELISA, CXCL8 Kit ELISA, il8 Kit ELISA, cxcl8a Kit ELISA, IL8 Kit ELISA, Cxcl8 Kit ELISA, LOC422654 Kit ELISA, cxcl8.L Kit ELISA
    Sujet
    Human IL-8 or Interleukin-8, also known as C-X-C Motif Chemokine 8 or Monocyte-derived Neutrophil Chemotactic Factor, is a 99 amino acid cytokine protein encoded by the IL8 gene located at locus 4q13-4q21 on chromosome 4. This gene is believed to play a role in the pathogenesis of bronchiolitis, a common respiratory tract disease caused by viral infection. The IL8 gene and other ten members of the CXC chemokine gene family form a chemokine gene cluster in a region mapped to chromosome 4q. IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T-cells, but not monocytes. Belonging to the Intercrine alpha (chemokine CxC) family, it is involved in neutrophil activation and is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively. Several N- terminal processed forms are produced by proteolytic cleavage after secretion from at least peripheral blood monocytes, leukocytes and endothelial cells. In general, IL-8(1-77) is referred to as interleukin-8 while IL-8(6-77) is the most prominent form. Source: Entrez Gene: IL8 interleukin 8 [Homo sapiens], Swiss-Prot: P10145
    ID gène
    3576
    UniProt
    P10145
    Pathways
    Signalisation TLR, Cellular Response to Molecule of Bacterial Origin, Regulation of G-Protein Coupled Receptor Protein Signaling, ER-Nucleus Signaling, Hepatitis C, Autophagy
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