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IGFBP3 ELISA 试剂盒

IGFBP3 适用: 小鼠 Colorimetric Sandwich ELISA 70-40000 pg/mL Cell Culture Supernatant, Plasma, Serum
产品编号 ABIN625402
发货至: 中国
  • 抗原 See all IGFBP3 ELISA试剂盒
    IGFBP3 (Insulin-Like Growth Factor Binding Protein 3 (IGFBP3))
    适用
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    小鼠
    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    70-40000 pg/mL
    最低检测浓度
    70 pg/mL
    应用范围
    ELISA
    原理
    Mouse IGFBP-3 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    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
    < 70 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
    Featured
    Discover our best selling IGFBP3 ELISA Kit
    Top Product
    Discover our top product IGFBP3 ELISA Kit
  • 应用备注
    Recommended Dilution for serum and plasma samples50 - 500 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 C (Item L) should be used for dilution of serum/plasma/culture supernatants. Suggested dilution for normal serum/plasma: 50-500 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 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 Assay Diluent C (Item L) into Item C vial to prepare a 100 ng/mL standard solution. Dissolve the powder thoroughly by a gentle mix. Add 200 µL IGFBP-3 standard from the vial of Item C, into a tube with 300 µL Assay Diluent C to prepare a 40,000 pg/mL standard solution. Pipette 400myl Assay Diluent C into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent C serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 200 µL standard + 300 µL 40,000 13,333 4,444 1,481 493.8 164.6 54.87 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 B (Item E) 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) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 500-fold with 1x Assay Diluent B (Item E). 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.
    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 C Mouse IGFBP-3 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 10 100 1,000 10,000 100,000
    Sensitivity: The minimum detectable dose of IGFBP-3 is typically less than 70 pg/mL.
    Recovery: Recovery was determined by spiking various levels of IGFBP-3 into normal Mouse serum, plasma (EDTA and Citrate) and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 85.83 73-94 Plasma 76.81 71-84 Cell culture media 105.5 93-112
    Linearity: Sample Type Serum Plasma Cell Culture Media (EDTA and Heparin) 1:2 Average % of Expected 104.0 106.2 91.21 Range ( %) 94-112 98-114 82-100 1:4 Average % of Expected 117.8 114.4 76.27 Range ( %) 106-125 104-123 70-85
    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
  • Obenauf, Zou, Ji, Vanharanta, Shu, Shi, Kong, Bosenberg, Wiesner, Rosen, Lo, Massagué: "Therapy-induced tumour secretomes promote resistance and tumour progression." in: Nature, Vol. 520, Issue 7547, pp. 368-72, (2015) (PubMed).

    Bickel, Feist, Müller, Quadbeck, Kekomäki: "Ornithinaemia. Another disorder of aminoacid metabolism associated with brain damage." in: German medical monthly, Vol. 14, Issue 3, pp. 101-5 (PubMed).

  • 抗原 See all IGFBP3 ELISA试剂盒
    IGFBP3 (Insulin-Like Growth Factor Binding Protein 3 (IGFBP3))
    别名
    IGFBP-3 (IGFBP3 产品)
    别名
    BP-53 ELISA Kit, IBP3 ELISA Kit, AI649005 ELISA Kit, IGFBP-3 ELISA Kit, IGgfbp3 ELISA Kit, IGF-BP3 ELISA Kit, zgc:91788 ELISA Kit, IGFBP3 ELISA Kit, igfbp3 ELISA Kit, insulin like growth factor binding protein 3 ELISA Kit, insulin-like growth factor binding protein 3 ELISA Kit, IGFBP3 ELISA Kit, Igfbp3 ELISA Kit, igfbp3 ELISA Kit, IGFBP-3 ELISA Kit, LOC100305016 ELISA Kit
    背景
    The Mouse IGFBP-3 (Insulin-like growth factor binding protein 3) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Mouse IGFBP-3 in serum, plasma ( citrate plasma samples are not recommended for use in this assay ) and cell culture supernatants. This assay employs an antibody specific for IGFBP-3 coated on a 96-well plate. Standards and samples are pipetted into the wells and IGFBP-3 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Mouse IGFBP-3 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 IGFBP-3 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
    16009
    UniProt
    P47878
    途径
    Myometrial Relaxation and Contraction, Regulation of Muscle Cell Differentiation, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Autophagy, Smooth Muscle Cell Migration, Growth Factor Binding
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