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MICA ELISA 试剂盒

MICA 适用: 人 Colorimetric Sandwich ELISA 20-10000 pg/mL Cell Culture Supernatant, Plasma, Serum
产品编号 ABIN625337
发货至: 中国
  • 抗原 See all MICA ELISA试剂盒
    MICA (MHC Class I Polypeptide-Related Sequence A (MICA))
    适用
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    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    20-10000 pg/mL
    最低检测浓度
    20 pg/mL
    应用范围
    ELISA
    原理
    Human MICA 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 following 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-6, 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 (OB), MCP-1, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, MMP-1, - 2, -3, -10, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    灵敏度
    20 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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    Discover our top product MICA ELISA Kit
  • 应用备注
    Recommended Dilution for serum and plasma samples2 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 (Item E) is used for dilution of serum/plasma/culture supernatants/urine. 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 100 ng/ml standard solution. Dissolve the powder thoroughly by a gentle mix. Add 50 µl MICA standard from the vial of tem C, into a tube with 450 µl 1x Assay Diluent Buffer to prepare a 10,000 pg/ml standard solution. Pipette 400 µl 1x Assay Diluent into each tube. Use the 10,000 pg/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Gently vortex to mix. 1x Assay Diluent serves as the zero standard (0 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 15,000-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 2 µl of HRP-Streptavidin concentrate into a tube with 198.0 µl 1x Assay Diluent to prepare a 100-fold diluted HRP-Streptavidin solution (do not store the diluted solution for next day use). Mix through and then pipette 80 µl of prepared 100-fold diluted solution into a tube with 12 ml 1x Assay Diluent to prepare a final 15,000 fold diluted HRP-Streptavidin solution.
    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 µl) 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 3. 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 3. 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.

    限制
    仅限研究用
  • 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
  • Shi, Li, Couturier, Yang, Guo, He, Lewis, Zhou: "Allele Specific Expression of MICA Variants in Human Fibroblasts Suggests a Pathogenic Mechanism." in: The open rheumatology journal, Vol. 9, pp. 60-4, (2015) (PubMed).

    Garcia-Chagollan, Jave-Suarez, Haramati, Sanchez-Hernandez, Aguilar-Lemarroy, Bueno-Topete, Pereira-Suarez, Fafutis-Morris, Cid-Arregui, del Toro-Arreola: "Substantial increase in the frequency of circulating CD4+NKG2D+ T cells in patients with cervical intraepithelial neoplasia grade 1." in: Journal of biomedical science, Vol. 20, pp. 60, (2013) (PubMed).

    Manno: "Measurement of the digit lengths and the anogenital distance in mice." in: Physiology & behavior, Vol. 93, Issue 1-2, pp. 364-8, (2008) (PubMed).

  • 抗原 See all MICA ELISA试剂盒
    MICA (MHC Class I Polypeptide-Related Sequence A (MICA))
    别名
    MICA (MICA 产品)
    别名
    MIC-A ELISA Kit, PERB11.1 ELISA Kit, MHC class I polypeptide-related sequence A ELISA Kit, MICA ELISA Kit
    背景
    The Human MICA (MHC class I Chain-related gene A) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human MICA in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human MICA coated on a 96-well plate. Standards and samples are pipetted into the wells and MICA present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human MICA 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 MICA 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
    100507436
    UniProt
    Q29983
    途径
    Activation of Innate immune Response, Transition Metal Ion Homeostasis, Human Leukocyte Antigen (HLA) in Adaptive Immune Response
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