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Galectin 3 Kit ELISA

LGALS3 Reactivité: Humain Colorimetric Sandwich ELISA Cell Culture Supernatant, Plasma, Serum
N° du produit ABIN955782
  • Antigène Voir toutes Galectin 3 (LGALS3) Kits ELISA
    Galectin 3 (LGALS3)
    Reactivité
    • 10
    • 9
    • 4
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Application
    ELISA
    Type d'échantillon
    Cell Culture Supernatant, Plasma, Serum
    Analytical Method
    Quantitative
    Specificité
    The interference of circulating factors of the immune system was evaluated by spiking these proteins at physiologically relevant concentrations into a Galectin-3 positive serum. There was no detectable cross reactivity.
    Sensibilité
    The limit of detection of human Galectin-3 is defined as the analyte concentration resulting in an absorption reading significantly higher than that of the dilution medium (mean plus two standard deviations) was determined to be 0.12 ng/mL (mean of 6 independent assays).
    Attributs du produit
    An anti-human Galectin-3 monoclonal coating antibody is adsorbed onto microwells. Human Galectin-3 present in the sample or calibrator binds to antibodies adsorbed to the microwells, a biotin-conjugated anti-human Galectin-3 antibody is added and binds to human Galectin-3 captured by the first antibody. Following incubation, unbound biotin-conjugated anti-human Galectin-3 is removed during a wash step. Streptavidin-HRP is added and binds to the biotin-conjugated anti-human Galectin-3 antibody. Following incubation, unbound streptavidin-HRP is removed during a wash step, and substrate solution reactive with HRP is added to the wells.
    Ingrédients
    1 aluminum pouch with a Microwell Plate coated with Monoclonal Antibody (mouse) to human Galectin-3
    1 vial (100 µL) Biotin-Conjugate anti-human Galectin-3 polyclonal antibody (rabbit)
    1 vial (150 µL) Streptavidin-HRP Concentrate
    2 vials human Galectin-3 Calibrator, lyophilized, 50 ng/mL upon reconstitution
    1 vial (5 mL) Assay Buffer Concentrate 20X (Phosphate-Buffered Saline with 1% Tween 20 and protein stabilizer)
    1 bottle (50 mL) Wash Buffer Concentrate 20X (Phosphate-Buffered Saline with 1% Tween 20)
    1 bottle (12 mL) Sample Diluent
    1 vial (15 mL) Substrate Solution (tetramethyl-benzidine)
    1 vial (12 mL) Stop Solution (1M Phosphoric acid)
    1 vial (0.4 mL) Blue Dye
    1 vial (0.4 mL) Red Dye
    1 vial (0.4 mL) Green Dye
    4 adhesive Plate Seals.
    Matériel non inclus
    5 mL and 10 mL graduated pipettes
    5 µL to 1,000 µL adjustable single channel micropipettes with disposable tips.
    50 µL to 300 µL adjustable multi-channel micropipette with disposable tips.
    Multi-channel micropipette reservoir.
    Beakers, flasks,
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  • Plaque
    Pre-coated
    Préparation des réactifs

    Buffer Concentrates should be brought to room temperature and should be diluted before starting the test procedure. If crystals have formed in the Buffer Concentrates, warm them gently until they have completely dissolved.

    a. Wash Buffer: Pour the entire contents (50 mL) of the Wash Buffer Concentrate into a clean 1,000 mL graduated cylinder. Bring the final volume to 1,000 mL with glass-distilled or de-ionized water. Mix gently to avoid foaming. The pH of the final solution should adjust to 7.4. Transfer to a clean wash bottle and store at 2°to 25°C. Please note that the Wash Buffer is stable for 30 days.

    b. Assay Buffer: Mix the contents of the bottle well. Add the contents of the Assay Buffer Concentrate (5.0 mL) to 95 mL distilled or de- ionized water and mix gently to avoid foaming. Store at 4°C. Please note that the Assay Buffer is stable for 30 days.

    c. Preparation of Biotin-Conjugate: Please note that the Biotin-Conjugate should be used within 30 minutes after dilution. The Biotin-Conjugate must be diluted 1:100 with Assay Buffer just prior to use in a clean plastic test tube

    d. Preparation of human Galectin-3 Calibrator: Reconstitute the human Galectin-3 Calibrator by addition of the volume of distilled water as stated on vial label. Mix or swirl gently to insure complete and homogeneous solubilization. (Concentration of reconstituted calibrator = 50 ng/mL)
    e. Preparation of Streptavidin-HRP: Please note that the Streptavidin-HRP should be used within 30 minutes after dilution. Make a 1:400 dilution of the Streptavidin-HRP Concentrate with Assay Buffer in a clean plastic tube as needed.

    f. Addition of Color Dyes (optional step): In order to help our customers to avoid any mistakes in pipetting, now offers a new tool that helps to monitor the addition of even very small volumes of a solution to the reaction well by giving distinctive colors to each step of the ELISA procedure. This procedure is optional, does not in any way interfere with the test results, and is designed to help the customer with the performance of the test, but can also be omitted.

    Préparation de l'échantillon

    Cell culture supernatants, human serum, plasma (EDTA) or other biological samples will be suitable for use in the assay. Remove the serum or plasma from the clot or red cells, respectively, as soon as possible after clotting and separation. Samples containing a visible precipitate must be clarified prior to use in the assay. Do not use grossly hemolyzed or lipemic specimens. Samples should be aliquoted and must be stored frozen at -20°C to avoid loss of bioactive Galectin-3. Addition of protease inhibitors may account for better stability of samples. If samples are to be run within 24 hours, they may be stored at 4°C. Avoid repeated freeze-thaw cycles. Prior to assay, the frozen sample should be brought to room temperature slowly and mixed gently. Pay attention to a possible Hook Effect due to high sample concentrations.

    Calcul des résultats

    Calculate the average absorbance values for each set of duplicate calibrators and samples. Duplicates should be within 20% of the mean.
    Create a calibration curve by plotting the mean absorbance for each calibrator concentration on the ordinate against the human Galectin-3 concentration on the abscissa. Draw a best-fit curve through the points of the graph (a 5- parameter curve fit is recommended).
    To determine the concentration of circulating human Galectin-3 for each sample, first find the mean absorbance value on the ordinate and extend a horizontal line to the calibration curve. At the point of intersection, extend a vertical line to the abscissa and read the corresponding human Galectin-3 concentration.
    For samples which have been diluted according to the instructions given in this manual (1:2) the concentration read from the calibration curve must be multiplied by the dilution factor (x2). Note: Calculation of samples with a concentration exceeding calibrator 1 may result in incorrect, low human Galectin-3 levels (Hook Effect). Such samples require further external predilution according to expected Galectin-3 values with Sample Diluent in order to precisely quantitate the actual human Galectin-3 level.
    It is suggested that each testing facility establishes a control sample of known human Galectin-3 concentration and runs this additional control with each assay. If the values obtained are not within the expected range of this control, the assay results may be invalid.

    Restrictions
    For Research Use only
  • Conseil sur la manipulation
    Since exact conditions may vary from assay to assay, a calibration curve must be established for every run.
    Bacterial or fungal contamination of either samples or reagents or cross-contamination between reagents may cause erroneous results.
    Stock
    4 °C
    Stockage commentaire
    Store kit reagents at 4°C. Immediately after use remaining reagents should be returned to cold storage (4°C). Expiration date of the kit and reagents is stated on labels. The expiration date of the kit components can only be guaranteed if the components are stored properly, and if, in case of repeated use of one component, the reagent is not contaminated by the first handling.
    Date de péremption
    The expiry date is stated on the label.
  • Antigène Voir toutes Galectin 3 (LGALS3) Kits ELISA
    Galectin 3 (LGALS3)
    Autre désignation
    Galectin-3 (LGALS3 Produits)
    Synonymes
    CBP 35 Kit ELISA, Gal-3 Kit ELISA, Galectin-3 Kit ELISA, leg3 Kit ELISA, galactoside-binding Kit ELISA, galectin-3 Kit ELISA, lectin Kit ELISA, soluble Kit ELISA, CBP35 Kit ELISA, GAL3 Kit ELISA, GALBP Kit ELISA, GALIG Kit ELISA, L31 Kit ELISA, LGALS2 Kit ELISA, MAC2 Kit ELISA, MGC52947 Kit ELISA, MGC75803 Kit ELISA, LGALS3 Kit ELISA, gal-3 Kit ELISA, GBP Kit ELISA, L-34 Kit ELISA, Mac-2 Kit ELISA, gal3 Kit ELISA, galectin 3 Kit ELISA, Galectin-3 Kit ELISA, avian beta-defensin 3 Kit ELISA, lectin, galactoside-binding, soluble, 3 L homeolog Kit ELISA, lectin, galactoside-binding, soluble, 3 Kit ELISA, galectin 2 Kit ELISA, lectin, galactose binding, soluble 3 Kit ELISA, LGALS3 Kit ELISA, leg3 Kit ELISA, AvBD3 Kit ELISA, lgals3.L Kit ELISA, lgals3 Kit ELISA, LGALS2 Kit ELISA, Lgals3 Kit ELISA
    Classe de substances
    Chemical
    Sujet
    Galectin-3 is a 26 kDa -galactoside-binding protein belonging to the Galectin family, which consists of more than ten members. Galectin-3 is composed of a carboxyl-terminal carbohydrate recognition domain (CRD) and amino-terminal tandem repeats. Galectin-3 normally distributes in epithelia of many organs and various inflammatory cells, including macrophages as well as dendritic cells and Kupfer cells. The expression of this lectin is up-regulated during inflammation, cell proliferation and cell differentiation and through transactivation by viral proteins. Its expression is also affected by neoplastic transformation: up-regulation is found in certain types of lymphomas, and thyroid carcinoma, while it is down regulated in other types of malignancies such as colon, breast, ovarian and uterine carcinomas. The expression of Galectin-3 has a strong correlation with the grade and malignant potential of primary brain tumors. Increased Galectin-3 levels have also been noted in human atherosclerotic lesions. These findings suggest that Galectin-3 expression is affected during these physiological and pathological responses. Galectin-3 has been shown to function through both intracellular and extracellular actions. It is a component of heterogeneous nuclear ribonuclear protein (hnRNP), a factor in pre-mRNA splicing and has been found to control cell cycle and prevent T-cell apoptosis through interaction with the Bcl-2 family members. On the other hand, this protein, which is secreted from monocytes/macrophages and epithelial cells has been demonstrated to function as an extracellular molecule in activating various types of cells such as monocytes/ macrophages, mast cells, neutrophils and lymphocytes. Galectin-3 has been shown to mediate cell-cell and cell- extracellular matrix interactions and acts as a novel chemoattractant for monocytes and macrophages.
    Pathways
    Signalisation RTK
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