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Decorin ELISA 试剂盒

DCN 适用: 小鼠 Colorimetric Sandwich ELISA 5-1000 pg/mL Cell Culture Supernatant, Plasma, Serum
产品编号 ABIN625382
发货至: 中国
  • 抗原 See all Decorin (DCN) ELISA试剂盒
    Decorin (DCN)
    适用
    • 13
    • 6
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    小鼠
    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    5-1000 pg/mL
    最低检测浓度
    5 pg/mL
    应用范围
    ELISA
    原理
    Mouse Decorin ELISA Kit for cell culture supernatants, Heparin and/or EDTA treated plasma, and serum samples. Citrate is not recommended.
    样品类型
    Serum, Plasma, Cell Culture Supernatant
    Analytical Method
    Quantitative
    特异性
    This ELISA kit shows no cross-reactivity with the following cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IFN- gamma, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-3 Rb, IL-4, IL-5, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LEPTIN(OB), LIX, L-Selectin, Lymphotactin, MCP-1, MCP- 5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, PF-4, PSelectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF RI, TNF RII, TPO, VCAM-1, 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 samples200 - 2,000 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, 1x Assay Diluent (Item E) should be used for dilution of serum/plasma/culture supernatants. Suggested dilution for normal serum/plasma: 200-2,000 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 (Item E) should be diluted 5-fold with deionized or distilled water before use.
      4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Assay Diluent (Item E) into Item C vial to prepare a 50 ng/mL standard solution. Dissolve the powder thoroughly by a gentle mix. Add 10 µL Decorin standard solution from the vial of Item C, into a tube with 490 µL 1x Assay Diluent to prepare a 1,000 pg/mL standard solution. Pipette 300myl 1x Assay Diluent into each tube. Use the 1,000 pg/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 10 µL standard + 490 µL 1,000 400 160 64 25.6 10.2 4.1 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/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 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 and used in step 4 of Part VI Assay Procedure.
      7. 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 300-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 40 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent to prepare a 300-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 Mouse Decorin concentration (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 Decorin is typically less than 5 pg/mL.
    Recovery: Recovery was determined by spiking various levels of Decorin into normal mouse serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 104.7 83-126 Plasma 93.91 74-119 Cell culture media 111.1 68-130
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 103.8 107.7 106.5 Range ( %) 96-112 100-116 101-112 1:4 Average % of Expected 114.0 106.0 106.1 Range ( %) 106-122 97-114 101-111
    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
  • 抗原 See all Decorin (DCN) ELISA试剂盒
    Decorin (DCN)
    别名
    Decorin (DCN 产品)
    别名
    CSCD ELISA Kit, DSPG2 ELISA Kit, PG40 ELISA Kit, PGII ELISA Kit, PGS2 ELISA Kit, SLRR1B ELISA Kit, DC ELISA Kit, mDcn ELISA Kit, cscd ELISA Kit, dspg2 ELISA Kit, pg40 ELISA Kit, pgii ELISA Kit, pgs2 ELISA Kit, slrr1b ELISA Kit, decorin ELISA Kit, decorin L homeolog ELISA Kit, DCN ELISA Kit, Dcn ELISA Kit, dcn.L ELISA Kit, dcn ELISA Kit
    基因ID
    13179
    UniProt
    P28654
    途径
    Glycosaminoglycan Metabolic Process
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