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FAS ELISA 试剂盒

FAS 适用: 人 Colorimetric Sandwich ELISA 5-2000 pg/mL Cell Culture Supernatant, Plasma, Serum
产品编号 ABIN624971
发货至: 中国
  • 抗原 See all FAS ELISA试剂盒
    FAS (TNF Receptor Superfamily, Member 6 (FAS))
    适用
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    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    5-2000 pg/mL
    最低检测浓度
    5 pg/mL
    应用范围
    ELISA
    原理
    Human Fas (TNFRSF6/Apo-1) ELISA Kit for cell culture supernatants, plasma, and serum samples.
    样品类型
    Plasma, Cell Culture Supernatant, Serum
    Analytical Method
    Quantitative
    特异性
    This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    灵敏度
    < 5 pg/mL
    产品特性
    • 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
    组件
    • 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
    试剂未包括
    • 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
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  • 应用备注
    Recommended Dilution for serum and plasma samples2 - 5 fold
    样本量
    100 μL
    板类型
    Pre-coated
    实验流程
    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.
    试剂准备
    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 supernantants and urine. Suggested dilution for normal serum/plasma: 2-5 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
      3. Assay Diluent B (Item B) should be diluted 5-fold with deionized or distilled water. 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.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 400myl Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (Assay Diluent B should be diluted 5-fold with deionized or distilled water, for cell culture medium and urine) into Item C vial to prepare 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL Fas standard from the vial of Item C, into a tube with 960 µL Assay Diluent A or 1x Assay Diluent B to prepare a 2,000 pg/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 pg/mL). 40 µL standard + 960 µL 200 µL 200 µL 200 µL 200 µL 200 µL 200 µL 2,000 666.7 222.2 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. 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.
      6. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 640-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 25 µL of HRP-Streptavidin concentrate into a tube with 16 ml 1x Assay Diluent B to prepare a final 640 fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    实验流程
    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.
    结果分析

    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 Fas concentrations (pg/mL) O D =4 50 n m 0.001 0.01 0.1 1 10 1 10 100 1,000 10,000 Assay Diluent B Human Fas concentrations (pg/mL) O D =4 50 n m 0.001 0.01 0.1 1 10 1 10 100 1,000 10,000
    Sensitivity: The minimum detectable dose of Fas is typically less than 5 pg/mL.
    Recovery: Recovery was determined by spiking various levels of recombinant human Fas into normal human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 84.46 78-102 Plasma 86.37 80-104 Cell culture media 91.24 82-105
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 83 84 87 Range ( %) 76-102 78-103 80-104 1:4 Average % of Expected 87 86 90 Range ( %) 80-104 79-104 83-105
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    实验精密度
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    限制
    仅限研究用
  • 注意事项
    Avoid repeated freeze-thaw cycles.
    储存条件
    -20 °C
    储存方法
    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.
    有效期
    6 months
  • Wu, Ding, Han, Arriens, Wei, Han, Pedroza, Jiang, Anolik, Petri, Sanz, Saxena, Mohan: "Antibody-Array-Based Proteomic Screening of Serum Markers in Systemic Lupus Erythematosus: A Discovery Study." in: Journal of proteome research, Vol. 15, Issue 7, pp. 2102-14, (2016) (PubMed).

    Richards, Timulak, Doherty, Sharry, Colla, Joyce, Hayes: "Internet-delivered treatment: its potential as a low-intensity community intervention for adults with symptoms of depression: protocol for a randomized controlled trial." in: BMC psychiatry, Vol. 14, pp. 147, (2014) (PubMed).

    Emmert-Streib, Abogunrin, de Matos Simoes, Duggan, Ruddock, Reid, Roddy, White, OKane, ORourke, Anderson, Nambirajan, Williamson: "Collectives of diagnostic biomarkers identify high-risk subpopulations of hematuria patients: exploiting heterogeneity in large-scale biomarker data." in: BMC medicine, Vol. 11, pp. 12, (2013) (PubMed).

    Abogunrin, OKane, Ruddock, Stevenson, Reid, OSullivan, Anderson, ORourke, Duggan, Lamont, Boyd, Hamilton, Nambirajan, Williamson: "The impact of biomarkers in multivariate algorithms for bladder cancer diagnosis in patients with hematuria." in: Cancer, Vol. 118, Issue 10, pp. 2641-50, (2012) (PubMed).

    Malard, Berenguer, Prat, Ruat, Steinmetz, Quemeneur: "Global gene expression profiling in human lung cells exposed to cobalt." in: BMC genomics, Vol. 8, pp. 147, (2007) (PubMed).

    Szabò, Gulbins, Apfel, Zhang, Barth, Busch, Schlottmann, Pongs, Lang: "Tyrosine phosphorylation-dependent suppression of a voltage-gated K+ channel in T lymphocytes upon Fas stimulation." in: The Journal of biological chemistry, Vol. 271, Issue 34, pp. 20465-9, (1996) (PubMed).

    Mapara, Bargou, Zugck, Döhner, Ustaoglu, Jonker, Krammer, Dörken: "APO-1 mediated apoptosis or proliferation in human chronic B lymphocytic leukemia: correlation with bcl-2 oncogene expression." in: European journal of immunology, Vol. 23, Issue 3, pp. 702-8, (1993) (PubMed).

  • 抗原 See all FAS ELISA试剂盒
    FAS (TNF Receptor Superfamily, Member 6 (FAS))
    别名
    Fas (FAS 产品)
    别名
    ALPS1A ELISA Kit, APO-1 ELISA Kit, APT1 ELISA Kit, CD95 ELISA Kit, FAS1 ELISA Kit, FASTM ELISA Kit, TNFRSF6 ELISA Kit, AI196731 ELISA Kit, APO1 ELISA Kit, TNFR6 ELISA Kit, Tnfrsf6 ELISA Kit, lpr ELISA Kit, Fas cell surface death receptor ELISA Kit, Fas (TNF receptor superfamily member 6) ELISA Kit, FAS ELISA Kit, Fas ELISA Kit, fas ELISA Kit
    背景
    Fas (APO-1 or CD95) is a cell-surface receptor that transduces apoptotic signals from Fas ligand (FasL). Fas and Fas Ligand (FasL) belong to the TNF superfamily and are type I and type II transmembrane proteins, respectively. Fas and FasL have been observed as soluble molecules in addition to their membraneassociated forms. Fas is expressed to a large extent on activated T and B lymphocytes, and on malignant lymphoid cells. The Human Fas ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human Fas in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human Fas coated on a 96-well plate. Standards and samples are pipetted into the wells and Fas present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human Fas 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 Fas 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%.
    基因ID
    355
    UniProt
    P25445
    途径
    p53 Pathway, Apoptosis, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity
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