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Notch1 ELISA 试剂盒

NOTCH1 适用: 人 Colorimetric Sandwich ELISA 79-5000 pg/mL Cell Lysate, Plasma, Serum
产品编号 ABIN1379982
发货至: 中国
  • 抗原 See all Notch1 (NOTCH1) ELISA试剂盒
    Notch1 (NOTCH1) (Notch 1 (NOTCH1))
    适用
    • 3
    • 3
    • 2
    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    79-5000 pg/mL
    最低检测浓度
    79 pg/mL
    应用范围
    ELISA
    原理
    The OmniKine? Human Notch-1 ELISA Kit contains the components necessary for quantitative determination of natural or recombinant Human Notch-1 concentrations within any experimental sample including cell lysates, serum and plasma. This particular immunoassay utilizes the quantitative technique of a "Sandwich" Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a "sandwich" format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on Human Notch-1 while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non-specific binding between proteins to other proteins or to the solid phase. After incubation and "sandwiching" of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a colorimetric reaction to ensue upon substrate addition. When the substrate TMB (3, 3', 5, 5'-Tetramethylbenzidine) is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution (2 N Sulfuric Acid) where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.
    品牌
    OmniKine™
    样品类型
    Cell Lysate, Serum, Plasma
    Analytical Method
    Quantitative
    特异性
    The Human Notch-1 ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human Notch-1 proteins.
    交叉反应 (详细)
    The Human Notch-1 ELISA is capable of recognizing both recombinant and naturally produced Human Notch-1 proteins.The antigens listed below were tested at 50 ng/mL and did not exhibit significant cross reactivity or interference. Human: DLL1, DNER, Jagged 1/Fc Chimera, Jagged 2/Fc Chimera, Notch-2/Fc Chimera, Notch-3/Fc Chimera Murine: Notch-1/Fc Chimera Rat: Notch-1/Fc Chimera, Notch-2/Fc Chimera
    产品特性
    The Human Notch-1 ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human Notch-1 proteins within the range of 79-5000 pg/mL.
    组件
    • Microstrips Coated w / Capture Antibody: 12 x 8-Well Microstrips
    • Protein Standard: Lyophilized (100 ng), Red container
    • Biotinylated Detection Antibody: Lyophilized, Yellow container
    • 400x Streptavidin-HRP: 30 μL, Blue container
    • Wash Buffer (10x): 50 mL, Clear containter
    • Assay Diluent: 50 mL, Clear container
    • Ready-to-Use Substrate: 12 mL, Brown container
    • Stop Solution: 12 mL, Clear container
    • Adhesive Plate Sealers: 4 Sheets
    • Technical Manual 1 Manual
    试剂未包括
    The following materials and/or equipment are NOT provided in this kit but are necessary to successfully conduct the experiment:
    Microplate reader able to measure absorbance at 450 nm (with correction wavelength set to 540 nm or 570 nm)
    Micropipettes with capability of measuring volumes ranging from 1 μl to 1 mL
    Deionized or sterile water
    Squirt bottle, manifold dispenser, multichannel pipette reservoir or automated microplate washer
    Graph paper or computer software capable of generating or displaying logarithmic functions
    Absorbent paper or vacuum aspirator
    Test tubes or microfuge tubes capable of storing ≥1 mL
    Bench
    top centrifuge (optional)
    Bench
    top vortex (optional)
    Orbital shaker (optional)
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  • 板类型
    Pre-coated
    实验流程
    This particular immunoassay utilizes the quantitative technique of a Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a sandwich format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on the Human Notch-1 cytokine while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non-specific binding between proteins to other proteins or to the solid phase. After incubation and sandwiching of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a colorimetric reaction to ensue upon substrate addition. When the substrate TMB (3, 3’, 5, 5’-Tetramethylbenzidine) is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution (2 N Sulfuric Acid) where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.
    样品制备

    If samples are to be used within 24 hours, aliquot and store at 4 °C. If samples are to be used over a long period of time, aliquot and store between -20 °C and -80 °C, depending on the duration of storage.
    Note: Samples containing a visible precipitate or pellet must be clarified prior to use in the assay.
    Caution: Avoid repeated freeze/thaw cycles to prevent loss of biological activity of proteins in experimental samples.

    • Cell Lysate and Supernatants:
      Remove large cell components via centrifugation and perform the assay. Cell lysates and supernatants require a dilution using Assay Diluent. A serial dilution may be performed to determine a suitable dilution factor for the sample. For future use of the sample, follow the sample storage guidelines stated above.
    • Serum:
      Allow samples to clot in a serum separator tube (SST) for 30 minutes. After sufficient clotting, centrifuge at 1000 x g for 15 minutes and remove serum from SST in preparation for the assay. Serum samples require at least a 1:50 dilution using Assay Diluent. For future use of the sample, follow the storage guidelines above.
    • Plasma:
      Use heparin, citrate or EDTA as an anticoagulant to gather plasma from original biological sample. After collection of the plasma, centrifuge for 15 minutes at 1000 x g. This step must be performed within 30 minutes of plasma collection. Plasma samples require at least a 1:50 dilution using Assay Diluent. Afterwards, perform the assay or for future use of the sample, follow the storage guidelines stated above.

    实验流程

    Note: If possible, all incubation steps should be performed on an orbital shaker to equilibrate solutions when added to the microplate wells. Also, all provided solutions should be at ambient temperature prior to use.
    Note: Avoid adding solutions into wells at an angle, always keep pipette tip perpendicular to plate bottom.

    Reconstitution of Provided Materials:

      1. Reconstitute the Biotin-Conjugated Detection Antibody in 67 µL of ddH₂O for a concentration of 180 µg/ml.
      2. Reconstitute the Protein Standard in 100 µL of ddH₂O for a concentration of 340 ng/ml.
      3. Dilute the 50 mL of 10x Wash Buffer in 450 mL of ddH2O for 500 mL of 1x Wash Buffer.
    Addition of Known Standard and Unknown Sample to Immunoassay:
      The OmniKine™ Human CD163 ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human CD163 proteins

    结果分析

    Generation of Standard Curve and Interpretation of Data
    1. Average the duplicate or triplicate readings for each standard, control and sample and subtract the average zero standard optical density.
    2. Generate a standard curve by using Microsoft Excel or other computer software capable of establishing a 4- Parameter Logistic (4-PL) curve fit. If using Excel or an alternative graphing tool, plot the average optical density values in absorbance units (y-axis) against the known standard concentrations in pg/ml (x-axis).Note: Only use the values in which a noticeable gradient can be established. Afterwards, generate a best fit curve or trend-line through the plotted points via regression analysis. Note: Shown on the next page is an example of typical data produced by analysis of the standard sample.

    限制
    仅限研究用
  • 注意事项
    Reagents provided in this kit may be harmful if ingested, inhaled or absorbed through the skin. Please carefully review the MSDS for each reagent before conducting the experiment.
    Stop Solution contains 2 N Sulfuric Acid (H2SO4) and is an extremely corrosive agent. Please wear proper eye, hand and face protection when handling this material. When the experiment is finished, be sure to rinse the plate with copious amounts of running water to dilute the Stop Solution prior to disposing the plate.
    注意事项
    This ELISA kit is intended for research purposes only, NOT diagnostic or clinical procedures of any kind.
    Materials included in this kit should NOT be used past the expiration date on the kit label.
    Reagents or substrates included in this kit should NOT be mixed or substituted with reagents or substrates from any other kits.
    Variations in pipetting technique, washing technique, operator laboratory technique, kit age, incubation time or temperature may cause differences in binding affinity of the materials provided.
    The assay is designed to eliminate interference and background by other cellular macromolecules or factors present within any biological samples. However, the possibility of background noise cannot be fully excluded until all factors have been tested using the assay kit.

    Reagents provided in this kit may be harmful if ingested, inhaled or absorbed through the skin. Please carefully review the MSDS for each reagent before conducting the experiment.
    Stop Solution contains 2 N Sulfuric Acid (H2SO4) and is an extremely corrosive agent. Please wear proper eye, hand and face protection when handling this material. When the experiment is finished, be sure to rinse the plate with copious amounts of running water to dilute the Stop Solution prior to disposing the plate.
    储存条件
    4 °C
    储存方法
    Note: If used frequently, reagents may be stored at 4 °C.
    • Unopened Kits: Store at 4 °C for 6 months.
    • Microstrips Coated w/ Capture Antibody, 400x Streptavidin-HRP Wash Buffer (10x), Assay Diluent Ready-to-Use Substrate, Stop Solution: 6 Months at 4 °C
    • Protein Standard, Biotinylated Detection Antibody: Lyophilized: 6 Months (if Reconstituted: 1 Month) at 4 °C
  • 抗原 See all Notch1 (NOTCH1) ELISA试剂盒
    Notch1 (NOTCH1) (Notch 1 (NOTCH1))
    别名
    Notch-1 (NOTCH1 产品)
    别名
    notch1 ELISA Kit, NOTCH1 ELISA Kit, hn1 ELISA Kit, tan1 ELISA Kit, notch ELISA Kit, xotch ELISA Kit, xnotch ELISA Kit, notch-1 ELISA Kit, xnotch1 ELISA Kit, TAN1 ELISA Kit, hN1 ELISA Kit, 9930111A19Rik ELISA Kit, Mis6 ELISA Kit, N1 ELISA Kit, Tan1 ELISA Kit, lin-12 ELISA Kit, NOTCH ELISA Kit, Xotch ELISA Kit, notch1-a ELISA Kit, notch 1 ELISA Kit, fibropellin-1 ELISA Kit, Notch homolog 1, translocation-associated (Drosophila) ELISA Kit, Notch homolog 1, translocation-associated ELISA Kit, notch 1 S homeolog ELISA Kit, notch1 ELISA Kit, LOC100631526 ELISA Kit, NOTCH1 ELISA Kit, Notch1 ELISA Kit, notch1.S ELISA Kit
    背景
    Notch-1 functions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. This receptor affects the implementation of differentiation, proliferation and apoptotic programs, and may be important for normal lymphocyte function. In altered form, Notch-1 may contribute to transformation or progression in some T-cell neoplasms. While it is involved in the maturation of both CD4+ and CD8+ cells in the thymus, it may also be important for follicular differentiation and possibly cell fate selection within the follicle. During cerebellar development, may function as a receptor for neuronal DNER and may be involved in the differentiation of Bergmann glia. Notch-1 forms a heterodimer of a C-terminal fragment N(TM) and an N-terminal fragment N(EC) which are probably linked by disulfide bonds. The heterodimer interacts with DNER, DTX1, DTX2 and RBPJ/RBPSUH in addition to MAML1, MAML2 and MAML3 which act as transcriptional co- activators for NOTCH1.Studies have shown that in fetal tissues the receptor is most abundant in spleen, brain stem and lung, and also presents in most adult tissues where it is found mainly in lymphoid tissues.Notch-1 is synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase in the trans- Golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and an N-terminal fragment N(EC). Following ligand binding, it is cleaved by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin dependent gamma-secretase to release a notch-derived peptide containing the intracellular domain (NICD) from the membrane. Defects in Notch-1 are a cause of bicuspid aortic valve (BAV), a common defect in the aortic valve in which two rather than three leaflets are present. It is often associated with aortic valve calcification and insufficiency. In extreme cases, the blood flow may be so restricted that the left ventricle fails to grow, resulting in hypoplastic left heart syndrome.
    途径
    Notch Signaling, Stem Cell Maintenance, Regulation of Muscle Cell Differentiation, Tube Formation, Skeletal Muscle Fiber Development
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