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TGFB1 Kit ELISA

TGFB1 Reactivité: Humain Colorimetric Sandwich ELISA 18 pg/mL - 4000 pg/mL Cell Culture Supernatant, Plasma, Serum
N° du produit ABIN625094
  • 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
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    18 pg/mL - 4000 pg/mL
    Seuil minimal de détection
    18 pg/mL
    Application
    ELISA
    Fonction
    Human TGF beta 1 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Type d'échantillon
    Plasma, Cell Culture Supernatant, Serum
    Analytical Method
    Quantitative
    Specificité
    This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human ANG, CD23, Eotaxin, GCSF, GM-CSF, GRO-alpha, GRO-beta, GRO-gamma, I-309, IFN-gamma, IL-1 alpha, IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL-7
    Sensibilité
    18 pg/mL
    Attributs du produit
    • 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
    Ingrédients
    • 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
    Matériel non inclus
    • 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
    Featured
    Discover our best selling TGFB1 Kit ELISA
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    Discover our top product TGFB1 Kit ELISA
  • Indications d'application
    Recommended Dilution for serum and plasma samples3 fold after treatment (see activation steps on page 6)
    Volume d'échantillon
    100 μL
    Plaque
    Pre-coated
    Protocole
    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.
    Préparation des réactifs
    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 (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine.
      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. Add 700 µL Assay Diluent A (Item D) (for serum/plasma) or 1x Assay Diluent B (Item E) (for cell culture supernatants/urine) to prepare a 300 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 120 µL TGF-beta1 standard from the vial of Item C, into a tube with 480 µL Assay Diluent A or 1x Assay Diluent B to prepare a 60 ng/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 ng/mL). 200 µL 120 µL standard +480 µL 200myl 200 µL 200 µL 200 µL 200 µL 60 20 6.667 2.222 0.741 0.247 0.0823 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/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) 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 500-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well. *Reagents to activate cell culture supernate/urine samples and Serum/Plasma samples 1 N HCl (100 ml) - Slowly add 8.33 mL of 12 N HCl into 91.67 ml deionized water. Mix bottle. 1.2 N NaOH/0.5 M HEPES (100 ml) - Slowly add 12 ml of 10 N NaOH into 75 mL deionized water. Mix bottle. Add 11.9 g HEPES. Mix through. Bring final volume to 100 mL with deionized water. 2.5 N Acetic Acid/10 M Urea (250 ml) - Add 150.2 g of Urea into 100 mL deionized water. Mix bottle until dissolved. Slowly add 35.9 mL of Glacial Acetic Acid. Mix through. Bring final volume to 250 ml with deionized water. 2.7 N NaOH/1 M HEPES (250 ml) - Add 67.5 ml of 10 N NaOH into 140 ml deionized water. Mix bottle. Add 59.5 g HEPES. Mix through. Bring final volume to 250 mL with deionized water. VI. TGF-beta1 SAMPLE ACTIVATION PROCEDURE To activate latent TGF-beta1 to the immunoreactive form, follow the activation procedure outlined below. Assay samples after neutralization (pH 7.0 - 7.6). Use polypropylene test tubes. Notes: Do not activate the kit standards. The kit standards contain active rhTGF-beta1. 1. Cell Culture Supernates/Urine Add 0.1 ml 1 N HCI into 0.5 mL cell culture supernate or urine. Mix tube thoroughly. Incubate for 10 minutes at room temperature. Neutralize the acidified sample by adding 0.1 ml 1.2 N NaOH/0.5 M HEPES (PH=7.0~7.6). Mix tube thoroughly. Assay immediately. The activated sample may be diluted with 1x Assay Diluent B (for cell culture supernatants/urine). The concentration read off the standard curve must be multiplied by the dilution factor.
      2. Serum/plasma Add 0.1 ml 2.5 N Acetic Acid/10 M Urea to 0.1 ml serum. Mix tube thoroughly. Incubate for 10 minutes at room temperature. Neutralize the acidified sample by adding 0.1 ml 2.7 N NaOH/1 M HEPES. Mix tube thoroughly. Assay immediately. The activated sample may be diluted with Assay Diluent A. The concentration read off the standard curve must be multiplied by the dilution factor.
    Procédure de l'essai
    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.
    Calcul des résultats

    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 TGF-beta-1 concentration (ng/mL) O D =4 50 n m 0.01 0.1 1 10 0 0.1 1 10 100 Assay Diluent B Human TGF-beta-1 concentration (ng/mL) O D =4 50 n m 0.01 0.1 1 10 0 0.1 1 10 100
    Sensitivity: The minimum detectable dose of TGF-beta1 is typically less than 80 pg/mL.
    Recovery: Recovery was determined by spiking various levels of human TGF-beta1 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 94.46 82-102 Plasma 95.78 93-103 Cell culture media 97.87 85-104
    Linearity: Sample Type Serum Plasma Cell culture media 1:2 Average % of Expected 92 95 95 Range ( %) 82-103 83-104 84-104 1:4 Average % of Expected 93 94 94 Range ( %) 83-105 84-105 83-104
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Précision du teste
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Restrictions
    For Research Use only
  • Conseil sur la manipulation
    Avoid repeated freeze-thaw cycles.
    Stock
    -20 °C
    Stockage commentaire
    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.
    Date de péremption
    6 months
  • Passanha, Geuens, LaPointe: "Cadherin-11 influences differentiation in human mesenchymal stem cells by regulating the extracellular matrix via the TGFβ1 pathway." dans: Stem cells (Dayton, Ohio), (2022) (PubMed).

    Prestigiacomo, Suter-Dick: "Nrf2 protects stellate cells from Smad-dependent cell activation." dans: PLoS ONE, Vol. 13, Issue 7, pp. e0201044, (2019) (PubMed).

    Xing, Xiao, Lu, Zhu, He, Huang, Lopez, Wong, Ju, Tian, Zhang, Xu, Wang, Li, Karin, Ren: "GFI1 downregulation promotes inflammation-linked metastasis of colorectal cancer." dans: Cell death and differentiation, Vol. 24, Issue 5, pp. 929-943, (2018) (PubMed).

    Jha, Mathur, Svedlund, Ye, Yeghiazarians, Healy: "Molecular weight and concentration of heparin in hyaluronic acid-based matrices modulates growth factor retention kinetics and stem cell fate." dans: Journal of controlled release : official journal of the Controlled Release Society, Vol. 209, pp. 308-16, (2016) (PubMed).

    Khoo, Nadarajan, Shim, Miswan, Zang, Possinger, Elstner: "Pretreatment of BMSCs with TZD solution decreases the proliferation rate of MCF‑7 cells by reducing FGF4 protein expression." dans: Molecular medicine reports, Vol. 13, Issue 4, pp. 3406-14, (2016) (PubMed).

    Kadam, Chuan: "Erratum to: Rectocutaneous fistula with transmigration of the suture: a rare delayed complication of vault fixation with the sacrospinous ligament." dans: International urogynecology journal, Vol. 27, Issue 3, pp. 505, (2016) (PubMed).

    Diaz-Gomez, Concheiro, Alvarez-Lorenzo, García-González: "Growth factors delivery from hybrid PCL-starch scaffolds processed using supercritical fluid technology." dans: Carbohydrate polymers, Vol. 142, pp. 282-92, (2016) (PubMed).

    Sun, Xia, Zhang, Liu, Shi: "P53 is required for Doxorubicin-induced apoptosis via the TGF-beta signaling pathway in osteosarcoma-derived cells." dans: American journal of cancer research, Vol. 6, Issue 1, pp. 114-25, (2016) (PubMed).

    Jha, Tharp, Ye, Santiago-Ortiz, Jackson, Stahl, Schaffer, Yeghiazarians, Healy: "Enhanced survival and engraftment of transplanted stem cells using growth factor sequestering hydrogels." dans: Biomaterials, Vol. 47, pp. 1-12, (2015) (PubMed).

    El-Haggar, Mostafa et al.: "Comparative clinical study between the effect of fenofibrate alone and its combination with pentoxifylline on biochemical parameters and liver stiffness in patients with non-alcoholic fatty liver ..." dans: Hepatology international, Vol. 9, Issue 3, pp. 471-9, (2015) (PubMed).

    Sowmya, Chennazhi, Arzate, Jayachandran, Nair, Jayakumar: "Periodontal Specific Differentiation of Dental Follicle Stem Cells into Osteoblast, Fibroblast, and Cementoblast." dans: Tissue engineering. Part C, Methods, (2015) (PubMed).

    Chen, Min, Wang, Leung, Shi, Zhou, Yu, Wang, An, Sha, Chen: "Pre-activation of mesenchymal stem cells with TNF-?, IL-1? and nitric oxide enhances its paracrine effects on radiation-induced intestinal injury." dans: Scientific reports, Vol. 5, pp. 8718, (2015) (PubMed).

    Shen, Lie, Miao, Yu, Lu, Feng, Li, Zu, Liu, Li: "Conditioned medium from umbilical cord mesenchymal stem cells induces migration and angiogenesis." dans: Molecular medicine reports, Vol. 12, Issue 1, pp. 20-30, (2015) (PubMed).

    Tang, Chen, Guo, Yang, Tao, Li, Miao, Feng, Chen, Zhu: "Minocycline Attenuates Neonatal Germinal-Matrix-Hemorrhage-Induced Neuroinflammation and Brain Edema by Activating Cannabinoid Receptor 2." dans: Molecular neurobiology, (2015) (PubMed).

    Yi, Yu, Zhang, Song, Jiang, Du, Wang: "Cathelicidin-BF suppresses intestinal inflammation by inhibiting the nuclear factor-κB signaling pathway and enhancing the phagocytosis of immune cells via STAT-1 in weanling piglets." dans: International immunopharmacology, Vol. 28, Issue 1, pp. 61-9, (2015) (PubMed).

    Koob, Lim, Zabek, Massee: "Cytokines in single layer amnion allografts compared to multilayer amnion/chorion allografts for wound healing." dans: Journal of biomedical materials research. Part B, Applied biomaterials, Vol. 103, Issue 5, pp. 1133-40, (2015) (PubMed).

    Koob, Lim, Massee, Zabek, Denozière: "Properties of dehydrated human amnion/chorion composite grafts: Implications for wound repair and soft tissue regeneration." dans: Journal of biomedical materials research. Part B, Applied biomaterials, Vol. 102, Issue 6, pp. 1353-62, (2014) (PubMed).

    Jun, Zhang, Yoon, Moon, Lee, Park, Kang, Lee, Kim, You: "Hypoxic conditioned medium from human amniotic fluid-derived mesenchymal stem cells accelerates skin wound healing through TGF-?/SMAD2 and PI3K/Akt pathways." dans: International journal of molecular sciences, Vol. 15, Issue 1, pp. 605-28, (2014) (PubMed).

    Chowdhury, Ahmed, Choudhuri, Sen, Hazra, Pal, Bhattacharya, Bahar: "Alteration of serum inflammatory cytokines in active pulmonary tuberculosis following anti-tuberculosis drug therapy." dans: Molecular immunology, Vol. 62, Issue 1, pp. 159-68, (2014) (PubMed).

    Gao, Ding, Xiao, Li, Chen, Zhou, Wang, Wu, Shi: "Anti-inflammatory and anti-apoptotic effect of combined treatment with methylprednisolone and amniotic membrane mesenchymal stem cells after spinal cord injury in rats." dans: Neurochemical research, Vol. 39, Issue 8, pp. 1544-52, (2014) (PubMed).

  • Antigène Voir toutes TGFB1 Kits ELISA
    TGFB1 (Transforming Growth Factor, beta 1 (TGFB1))
    Autre désignation
    TGF-beta 1 (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
    Transforming Growth Factor Beta (TGF-beta) is a stable, multifunctional polypeptide growth factor. TGF-beta exists in at least five isoforms, known as TGF-beta1, TGF-beta2, TGF-beta3, TGF-beta4, TGF-beta5. Their amino acid sequences display homologies on the order of 70-80%. The various TGF-beta isotypes share many biological activities and their actions on cells are qualitatively similar in most cases although there are a few examples of distinct activities. TGF-beta1 is the prevalent form and is found almost ubiquitously while the other isoforms are expressed in a more limited spectrum of cells and tissues. It is normally secreted as an inactive, or latent, complex. The Human TGF-beta1 ELISA (Enzyme-Linked mmunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human TGF-beta1 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human TGF-beta1 coated on a 96-well plate. Standards and samples are pipetted into the wells and TGF-beta1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human TGF-beta1 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 TGF-beta1 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%.
    ID gène
    7040
    UniProt
    P01137
    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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