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Insulin ELISA 试剂盒

INS 适用: 人 Colorimetric Sandwich ELISA 4-300 μIU/mL Cell Culture Supernatant, Plasma, Serum
产品编号 ABIN1980088
发货至: 中国
  • 抗原 See all Insulin (INS) ELISA试剂盒
    Insulin (INS)
    适用
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    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    4-300 μIU/mL
    最低检测浓度
    4 μIU/mL
    应用范围
    ELISA
    原理
    Human Insulin 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 the following cytokines tested: human BDNF, BLC, ENA-78, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), 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, TNF-alpha, TNF-beta, TPO, VEGF.
    灵敏度
    4 μIU/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 INS ELISA Kit
    Top Product
    Discover our top product INS ELISA Kit
  • 应用备注
    Recommended Dilution for serum and plasma samples2 - 10 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 supernatants and urine. Suggested dilution for normal serum/plasma: 2-10 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 should be diluted 5-fold with deionized or distilled water before use.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture supernates/urine) into Item C vial to prepare a 1,400 µIU/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 150 µL Insulin standard (1,400 µIU/mL) from the vial of Item C, into a tube with 550 µL Assay Diluent A or 1x Assay Diluent B to prepare a 300 µIU/mL standard solution. Pipette 300 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the 300 µIU/mL 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 µIU/mL). 300 µL 150 µL standard + 550 µL 300myl 300 µL 300 µL 300 µL 300 µL 300 150 75 37.5 18.75 9.38 4.69 0 µIU/mL µIU/mL µIU/mL µIU/mL µIU/mL µIU/mL µIU/mL µIU/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) and pipette up and down to mix gently 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 30 µL of HRP-Streptavidin concentrate into a tube with 15 ml 1x Assay Diluent B to prepare a final 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 A Human Insulin concentration (myIU/mL) O D =4 50 n m 0.01 0.1 1 10 1 10 100 1000 Assay Diluent B Human Insulin concentration (myIU/mL) O D =4 50 n m 0.01 0.1 1 10 1 10 100 1000
    Sensitivity: The minimum detectable dose of Insulin is typically less than 4 µIU/mL.
    Recovery: Recovery was determined by spiking various levels of human Insulin into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 91.40 83-102 Plasma 99.03 73-128 Cell culture media 76.16 68-88
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 100.2 109.5 86.69 Range ( %) 90-108 102-128 72-103 1:4 Average % of Expected 122.1 131.7 82.43 Range ( %) 112-135 121-140 68-90
    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
  • Jørgensen, Nellemann, Wohlfahrt-Veje, Jensen, Main, Andersen: "Interaction between paraoxonase 1 polymorphism and prenatal pesticide exposure on metabolic markers in children using a multiplex approach." in: Reproductive toxicology (Elmsford, N.Y.), Vol. 51, pp. 22-30, (2015) (PubMed).

    Townsend, Hoffman, Gonzalez, Jajtner, Boone, Robinson, Mangine, Wells, Fragala, Fukuda, Stout: "Effects of ?-Hydroxy-?-methylbutyrate Free Acid Ingestion and Resistance Exercise on the Acute Endocrine Response." in: International journal of endocrinology, Vol. 2015, pp. 856708, (2015) (PubMed).

    Leisegang, Bouic, Menkveld, Henkel: "Obesity is associated with increased seminal insulin and leptin alongside reduced fertility parameters in a controlled male cohort." in: Reproductive biology and endocrinology : RB&E, Vol. 12, pp. 34, (2014) (PubMed).

    Monteiro, Mota, Silveira, Cayres, Antunes, Fernandes, Freitas: "Morphological and metabolic determinants of nonalcoholic fatty liver disease in obese youth: a pilot study." in: BMC research notes, Vol. 6, pp. 89, (2013) (PubMed).

    Andersen, Wohlfahrt-Veje, Dalgård, Christiansen, Main, Nellemann, Murata, Jensen, Skakkebæk, Grandjean: "Paraoxonase 1 polymorphism and prenatal pesticide exposure associated with adverse cardiovascular risk profiles at school age." in: PLoS ONE, Vol. 7, Issue 5, pp. e36830, (2012) (PubMed).

    Kumar, Prakash, Chhabra, Singla, Madan, Gupta, Panda, Khanal, Acharya: "Association of pro-inflammatory cytokines, adipokines & oxidative stress with insulin resistance & non-alcoholic fatty liver disease." in: The Indian journal of medical research, Vol. 136, Issue 2, pp. 229-36, (2012) (PubMed).

    Kerem, Ferahkose, Yilmaz, Pasaoglu, Ofluoglu, Bedirli, Salman, Sahin, Akin: "Adipokines and ghrelin in gastric cancer cachexia." in: World journal of gastroenterology : WJG, Vol. 14, Issue 23, pp. 3633-41, (2008) (PubMed).

  • 抗原 See all Insulin (INS) ELISA试剂盒
    Insulin (INS)
    别名
    Insulin (INS 产品)
    别名
    IDDM2 ELISA Kit, ILPR ELISA Kit, IRDN ELISA Kit, MODY10 ELISA Kit, ins1 ELISA Kit, xins ELISA Kit, ins1-a ELISA Kit, Insulin ELISA Kit, AA986540 ELISA Kit, Ins-2 ELISA Kit, InsII ELISA Kit, Mody ELISA Kit, Mody4 ELISA Kit, proinsulin ELISA Kit, zgc:109842 ELISA Kit, igf2-A ELISA Kit, ins ELISA Kit, ins-a ELISA Kit, ins-b ELISA Kit, insulin ELISA Kit, insulin precursor ELISA Kit, insulin II ELISA Kit, preproinsulin ELISA Kit, insulin L homeolog ELISA Kit, insulin S homeolog ELISA Kit, INS ELISA Kit, INS-IGF2 ELISA Kit, ins ELISA Kit, Ins ELISA Kit, PIN ELISA Kit, Ins2 ELISA Kit, ins.L ELISA Kit, ins.S ELISA Kit
    背景
    The Human Insulin ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human Insulin and Proinsulin in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human Insulin coated on a 96-well plate. Standards and samples are pipetted into the wells and Insulin present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human Insulin 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 Insulin 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
    3630
    UniProt
    P01308
    途径
    NF-kappaB Signaling, RTK signaling, Positive Regulation of Peptide Hormone Secretion, Peptide Hormone Metabolism, Hormone Activity, Carbohydrate Homeostasis, ER-Nucleus Signaling, Regulation of Carbohydrate Metabolic Process, Feeding Behaviour, Autophagy, Negative Regulation of intrinsic apoptotic Signaling, Brown Fat Cell Differentiation, Positive Regulation of fat Cell Differentiation
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