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TGFB1 Kit ELISA

TGFB1 Reactivité: Porc Colorimetric Sandwich ELISA 15.6-1000 pg/mL Cell Culture Supernatant, Plasma, Serum
N° du produit ABIN579236
  • Antigène Voir toutes TGFB1 Kits ELISA
    TGFB1 (Transforming Growth Factor, beta 1 (TGFB1))
    Reactivité
    • 12
    • 8
    • 7
    • 6
    • 5
    • 4
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 1
    • 1
    Porc
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    15.6-1000 pg/mL
    Seuil minimal de détection
    15.6 pg/mL
    Application
    ELISA
    Fonction
    This immunoassay kit allows for the specific measurement of porcine TGFbeta1 concentrations in cell culture supernates, serum, and plasma.
    Type d'échantillon
    Cell Culture Supernatant, Serum, Plasma
    Analytical Method
    Quantitative
    Specificité
    This assay recognizes recombinant and natural porcine TGFbeta1.
    Réactivité croisée (Details)
    No significant cross-reactivity or interference was observed.
    Attributs du produit
    Sus scrofa,Pig,Transforming growth factor beta-1,TGF-beta-1,TGFB1
    Ingrédients
    Reagent (Quantity ): Assay plate (1), Standard (2), 2 Sample Diluent (1 × 20ml), Assay Diluent A (1x10ml), Assay Diluent B (1x10ml), Detection Reagent A (1 × 120μl), Detection Reagent B (1 × 120μl), Wash Buffer (25 x concentrate) (1 × 30ml), Substrate (1x10ml), Stop Solution (1 x 10ml)
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  • Volume d'échantillon
    100 μL
    Plaque
    Pre-coated
    Protocole
    This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for TGFbeta1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TGFbeta1 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for TGFbeta1 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TGFbeta1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
    Préparation des réactifs

    Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 500 mL of Wash Buffer. Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 1000 pg/mL. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions. The undiluted standard serves as the high standard (1000 pg/mL). The Sample Diluent serves as the zero standard (0 pg/mL). Detection Reagent A and B - Dilute to the working concentration specified on the vial label using Assay Diluent A and B (1:100), respectively.
    3.

    Prélèvement de l'échantillon
    Cell culture supernates - Remove particulates by centrifugation and assay immediately or aliquot and store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles. Serum - Use a serum separator tube (SST) and allow samples to clot for 30 minutes before centrifugation for 15 minutes at approximately 1000 x g. Remove serum and assay immediately or aliquot and store samples at -20 °C. Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 x g at 2 - 8 °C within 30 minutes of collection. Store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles. Note: Citrate plasma has not been validated for use in this assay.
    Procédure de l'essai

    Allow all reagents to reach room temperature. Arrange and label required number of strips.
    1. Prepare all reagents, working standards and samples as directed in the previous sections.
    2. Add 100 uL of Standard, Control, or sample* per well. Cover with the adhesive strip. Incubate for 2 hours at 37 °C.
    3. Remove the liquid of each well, don’t wash.
    4. Add 100 uL of Detection Reagent A to each well. Incubate for 1 hour at 37°C. Detection Reagent A may appear cloudy. Warm to room temperature and mix gently until solution appears uniform.
    5. Aspirate each well and wash, repeating the process three times for a total of three washes. Wash by filling each well with Wash Buffer (350 uL) using a squirt bottle, multi-channel pipette, manifold dispenser 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.
    6. Add 100 uL of Detection Reagent B to each well. Cover with a new adhesive strip.Incubate for 1 hours at 37 °C.
    7. Repeat the aspiration/wash as in step
    5. 8. Add 90 uL of Substrate Solution to each well. Incubate for 30 minutes at room temperature. Protect from light.
    9. Add 50 uL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
    10. Determine the optical density of each well within 30 minutes, using a microplate reader set to 450 nm.
    Important Note:1. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
    2. It is recommended that no more than 32 wells be used for each assay run if manual pipetting is used since pipetting of all standards, specimens and controls should be completed within 5 minutes. A full plate of 96 wells may be used if automated pipetting is available.
    3. Duplication of all standards and specimens, although not required, is recommended.
    4. When mixing or reconstituting protein solutions, always avoid foaming.
    5. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.
    6. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary.

    Calcul des résultats

    4 Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the TGFbeta1 concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

    Restrictions
    For Research Use only
  • Conseil sur la manipulation
    1. The kit should not be used beyond the expiration date on the kit label.
    2. Do not mix or substitute reagents with those from other lots or sources.
    3. If samples generate values higher than the highest standard, further dilute the samples with the Assay Diluent and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.
    4. This assay is designed to eliminate interference by soluble receptors, ligands, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.
    Stock
    4 °C/-20 °C
    Stockage commentaire
    The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 °C upon being received. The other reagents can be stored at 4 °C.
  • Antigène Voir toutes TGFB1 Kits ELISA
    TGFB1 (Transforming Growth Factor, beta 1 (TGFB1))
    Autre désignation
    TGFB1 (TGFB1 Produits)
    Synonymes
    CED Kit ELISA, DPD1 Kit ELISA, LAP Kit ELISA, TGFB Kit ELISA, TGFbeta Kit ELISA, TGF-beta Kit ELISA, TGF-BETA-1 Kit ELISA, TGF-beta5 Kit ELISA, ced Kit ELISA, dpd1 Kit ELISA, lap Kit ELISA, tgf-beta Kit ELISA, tgfb Kit ELISA, tgfb5 Kit ELISA, tgfbeta Kit ELISA, TGF-beta1 Kit ELISA, TGFbeta1 Kit ELISA, Tgfb Kit ELISA, Tgfb-1 Kit ELISA, ai39657 Kit ELISA, tgfb1 Kit ELISA, wu:fb13a07 Kit ELISA, xx:ai39657 Kit ELISA, TGFB1 Kit ELISA, csd Kit ELISA, cdb1 Kit ELISA, cdg2 Kit ELISA, csd1 Kit ELISA, csd2 Kit ELISA, csd3 Kit ELISA, ebmd Kit ELISA, lcd1 Kit ELISA, bigh3 Kit ELISA, cdgg1 Kit ELISA, betaig-h3 Kit ELISA, TGFB4 Kit ELISA, transforming growth factor beta 1 Kit ELISA, transforming growth factor beta-1 Kit ELISA, transforming growth factor beta 1 L homeolog Kit ELISA, transforming growth factor, beta 1 Kit ELISA, transforming growth factor, beta 1a Kit ELISA, transforming growth factor beta induced L homeolog Kit ELISA, TGFB1 Kit ELISA, Tgfb1 Kit ELISA, tgfb1.L Kit ELISA, tgfb1a Kit ELISA, tgfbi.L Kit ELISA
    Sujet
    TGF-beta is capable of producing a variety of effects and virtually all cell types respond to this factor in some way. The inappropriate presence of active TGF-beta1 has been implicated in a variety of pathological conditions Because of the necessity for regulating its activity tightly, TGF-beta is secreted by cells in the form of an inactive complex. This complex consists of TGF-beta1 associated non-covalently with a protein designated the latency associated peptide (LAP). TGF-beta 1 and LAP represent components of a pro-peptide that is cleaved in a post-golgi compartment prior to secretion. LAP and TGF-beta1 each consist of a disulfide-linked homodimer and the association of these two components renders TGF-beta1 inactive and inaccessible to anti-TGF-beta antibodies. In many instances in vivo, associated LAP and TGF-beta, called the small latent TGF-beta complex, are bound to an additional protein designated the latent TGF-beta1 binding protein (LTBP), forming the large latent TGF-beta1 complex. The mechanisms by which active TGF-beta1 is released from these latent complexes represent important control steps for the regulation of the numerous effects of TGF-beta1. Despite considerable study, these mechanisms are not well understood.
    ID gène
    2955
    Pathways
    EGFR Signaling Pathway, Dopaminergic Neurogenesis, Cellular Response to Molecule of Bacterial Origin, Glycosaminoglycan Metabolic Process, Regulation of Leukocyte Mediated Immunity, Regulation of Muscle Cell Differentiation, Positive Regulation of Immune Effector Process, Cell-Cell Junction Organization, Production of Molecular Mediator of Immune Response, Ribonucleoside Biosynthetic Process, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Protein targeting to Nucleus, Autophagy, Cancer Immune Checkpoints
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