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ERK2 ELISA 试剂盒

MAPK1 适用: 人 Colorimetric Sandwich ELISA 0.156-10 ng/mL Plasma, Serum
产品编号 ABIN455886
发货至: 中国
  • 抗原 See all ERK2 (MAPK1) ELISA试剂盒
    ERK2 (MAPK1) (Mitogen-Activated Protein Kinase 1 (MAPK1))
    适用
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    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    0.156-10 ng/mL
    最低检测浓度
    0.156 ng/mL
    应用范围
    ELISA
    原理
    This immunoassay kit allows for the specific measurement of human MAPK concentrations in serum and plasma.
    样品类型
    Serum, Plasma
    Analytical Method
    Quantitative
    特异性
    This assay recognizes recombinant and natural human MAPK.
    交叉反应 (详细)
    No significant cross-reactivity or interference was observed.
    产品特性
    Homo sapiens,Human,Mitogen-activated protein kinase 1,MAP kinase 1,MAPK 1,ERT1,Extracellular signal-regulated kinase 2,ERK-2,MAP kinase isoform p42,p42-MAPK,Mitogen-activated protein kinase 2,MAP
    组件
    Reagent (Quantity): Assay plate (1), Standard (2), Sample Diluent (1x20ml), Assay Diluent A (1x10ml), Assay Diluent B (1x10ml), Detection Reagent A 1×120µl Detection Reagent B 1×120µl Wash Buffer(25 x concentrate) (1x30ml), Substrate (1x10ml), Stop Solution (1x10ml)
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  • 样本量
    100 μL
    板类型
    Pre-coated
    实验流程
    This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for MAPK has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MAPK present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for MAPK is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of MAPK bound in the initial step. The color development is stopped and the intensity of the color is measured.
    试剂准备

    Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 500 mL of Wash Buffer. Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 10000 pg/mL. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions. The undiluted standard serves as the high standard (10000 pg/mL). The Sample Diluent serves as the zero standard (0 pg/mL). Detection Reagent A and B - Dilute to the working concentration specified on the vial label using Assay Diluent A and B (1:100), respectively.

    样品收集
    Serum - Use a serum separator tube (SST) and allow samples to clot for 30 minutes before centrifugation for 15 minutes at approximately 1000 x g. Remove serum and assay immediately or aliquot and store samples at -20 °C. Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 x g at 2 - 8 °C within 30 minutes of collection. Store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles. Note: Citrate plasma has not been validated for use in this assay.
    实验流程

    Allow all reagents to reach room temperature. Arrange and label required number of strips.
    1. Prepare all reagents, working standards and samples as directed in the previous sections.
    2. Add 100 uL of Standard, Control, or sample per well. Cover with the adhesive strip. Incubate for 2 hours at 37 °C.
    3. Remove the liquid of each well, don’t wash.
    4. Add 100 uL of Detection Reagent A to each well. Incubate for 1 hour at 37°C. Detection Reagent A may appear cloudy. Warm to room temperature and mix gently until solution appears uniform.
    5. Aspirate each well and wash, repeating the process three times for a total of three washes. Wash 3 by filling each well with Wash Buffer (350 uL) using a squirt bottle, multi-channel pipette, manifold dispenser 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.
    6. Add 100 uL of Detection Reagent B to each well. Cover with a new adhesive strip.Incubate for 1 hours at 37 °C.
    7. Repeat the aspiration/wash as in step
    5. 8. Add 90 uL of Substrate Solution to each well. Incubate for 30 minutes at room temperature. Protect from light.
    9. Add 50 uL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
    10. Determine the optical density of each well within 30 minutes, using a microplate reader set to 450 nm.
    Important Note:
    1. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
    2. It is recommended that no more than 32 wells be used for each assay run if manual pipetting is used since pipetting of all standards, specimens and controls should be completed within 5 minutes. A full plate of 96 wells may be used if automated pipetting is available.
    3. Duplication of all standards and specimens, although not required, is recommended.
    4. When mixing or reconstituting protein solutions, always avoid foaming.
    5. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.
    6. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary.

    结果分析

    Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the MAPK IGM concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

    限制
    仅限研究用
  • 注意事项
    1. The kit should not be used beyond the expiration date on the kit label.
    2. Do not mix or substitute reagents with those from other lots or sources.
    3. If samples generate values higher than the highest standard, further dilute the samples with the Assay Diluent and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.
    4. This assay is designed to eliminate interference by soluble receptors, ligands, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.
    储存条件
    4 °C/-20 °C
    储存方法
    The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 °C upon being received. The other reagents can be stored at 4 °C.
  • 抗原 See all ERK2 (MAPK1) ELISA试剂盒
    ERK2 (MAPK1) (Mitogen-Activated Protein Kinase 1 (MAPK1))
    别名
    MAPK1 (MAPK1 产品)
    别名
    ERK-1 ELISA Kit, ERK1 ELISA Kit, ERT2 ELISA Kit, HS44KDAP ELISA Kit, HUMKER1A ELISA Kit, P44ERK1 ELISA Kit, P44MAPK ELISA Kit, PRKM3 ELISA Kit, p44-ERK1 ELISA Kit, p44-MAPK ELISA Kit, Erk-1 ELISA Kit, Erk1 ELISA Kit, Ert2 ELISA Kit, Esrk1 ELISA Kit, Mnk1 ELISA Kit, Mtap2k ELISA Kit, Prkm3 ELISA Kit, p44 ELISA Kit, p44erk1 ELISA Kit, p44mapk ELISA Kit, ERK ELISA Kit, ERK2 ELISA Kit, ERT1 ELISA Kit, MAPK2 ELISA Kit, P42MAPK ELISA Kit, PRKM1 ELISA Kit, PRKM2 ELISA Kit, p38 ELISA Kit, p40 ELISA Kit, p41 ELISA Kit, p41mapk ELISA Kit, 9030612K14Rik ELISA Kit, AA407128 ELISA Kit, AU018647 ELISA Kit, C78273 ELISA Kit, Erk2 ELISA Kit, Prkm1 ELISA Kit, p42mapk ELISA Kit, zERK2 ELISA Kit, MAPK1 ELISA Kit, Xp42 ELISA Kit, erk ELISA Kit, erk2 ELISA Kit, ert1 ELISA Kit, mapk ELISA Kit, mapk1b ELISA Kit, mapk2 ELISA Kit, mpk1 ELISA Kit, prkm1 ELISA Kit, prkm2 ELISA Kit, mapk1 ELISA Kit, mapk1a ELISA Kit, xp42 ELISA Kit, ATMPK1 ELISA Kit, F14N23.9 ELISA Kit, F14N23_9 ELISA Kit, MITOGEN-ACTIVATED PROTEIN KINASE ELISA Kit, mitogen-activated protein kinase 1 ELISA Kit, MNK1 ELISA Kit, ATMPK2 ELISA Kit, MITOGEN-ACTIVATED PROTEIN KINASE HOMOLOG 2 ELISA Kit, T30E16.13 ELISA Kit, T30E16_13 ELISA Kit, mitogen-activated protein kinase homolog 2 ELISA Kit, mitogen-activated protein kinase 3 ELISA Kit, mitogen-activated protein kinase 1 ELISA Kit, mitogen activated protein kinase 1 ELISA Kit, mitogen-activated protein kinase 1 L homeolog ELISA Kit, mitogen-activated protein kinase 1 S homeolog ELISA Kit, mitogen activated protein kinase 3 ELISA Kit, mitogen-activated protein kinase homolog 2 ELISA Kit, MAPK3 ELISA Kit, Mapk3 ELISA Kit, MAPK1 ELISA Kit, Mapk1 ELISA Kit, mapk1 ELISA Kit, mapk1.L ELISA Kit, mapk1.S ELISA Kit, PfMAP1 ELISA Kit, MPK1 ELISA Kit, PVX_084965 ELISA Kit, LOC9328617 ELISA Kit, Tsp_01601 ELISA Kit, MPK2 ELISA Kit
    背景
    MAP kinase (MAPK), also known as a CDC-2-related protein kinase or CSBP (cytokine suppressive anti-inflammatory drug binding protein) is the human homolog of the Saccharomyces cerevisiae gene Hog1, which is a MAPK required for growth under highosmolarity conditions. MAPK is activated in response to a variety of extracellular stimuli including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), anisomycin, UV light and growth factors.
    途径
    MAPK Pathway, RTK signaling, Apoptosis, Interferon-gamma Pathway, Fc-epsilon Receptor Signaling Pathway, Response to Growth Hormone Stimulus, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Hepatitis C, Protein targeting to Nucleus, Toll-Like Receptors Cascades, Monocarboxylic Acid Catabolic Process, Autophagy, G-protein mediated Events, Signaling Events mediated by VEGFR1 and VEGFR2, Signaling of Hepatocyte Growth Factor Receptor, VEGFR1 Specific Signals, BCR Signaling, S100 Proteins
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