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CCL2 ELISA 试剂盒

CCL2 适用: 小鼠 Colorimetric Sandwich ELISA Cell Lysate, Tissue Lysate
产品编号 ABIN625160
发货至: 中国
  • 抗原 See all CCL2 ELISA试剂盒
    CCL2 (Chemokine (C-C Motif) Ligand 2 (CCL2))
    适用
    • 12
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    小鼠
    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    应用范围
    ELISA
    原理
    Mouse MCP-1 (CCL2) ELISA Kit for cell and tissue lysate samples.
    样品类型
    Tissue Lysate, Cell Lysate
    Analytical Method
    Quantitative
    特异性
    The antibody pair provided in this kit recognizes mouse MCP-1 / CCL2.
    灵敏度
    3 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
    • Cell lysate buffer
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  • 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: Tissue lysate and cell lysate sample should be diluted at least 5-folds with 1x Sample Diluent Buffer.
      3. Sample Diluent Buffer (Item D) and Assay Diluent (Item E) should be diluted 5-folds with deionized or distilled water before use.
      4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Sample Diluent Buffer (Item D, Sample Diluent Buffer should be diluted 5-fold with deionized or distilled water before use) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL MCP-1 standard from the vial of Item C, into a tube with 960 µL Sample Diluent Buffer to prepare a 2,000 pg/mL stock standard solution. Pipette 400 µL 1x Sample Diluent Buffer into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Sample Diluent Buffer serves as the zero standard (0 pg/mL). 200 µL 200 µL 200 µL 200myl 200 µL 200 µL 40 µL standard + 960 µL 2,000 666.7 222.2 74.07 24.69 8.23 2.74 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 Diuent 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-folds with 1x Assay Diuent and used in step 4 of Part VI Assay Procedure.
      7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) before use. HRP-Streptavidin concentrate should be diluted 400-folds with 1x Assay Diuent. For example: Add 30 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent to prepare a 400-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
      8. Cell lysate buffer should be diluted 2-fold with deionized or distilled water (for cell lysate and tissue lysate).
    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 Mouse MCP-1concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 (n m ) 0.01 0.1 1 10
    Sensitivity: The minimum detectable dose of MCP-1 is typically less than 3 pg/mL.
    Recovery: Recovery was determined by spiking various levels of mouse MCP-1 into mouse tissue lysate and cell lysate. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Tissue lysate 85.89 78-101 Cell lysate 87.56 80-102
    Linearity: Sample Type Tissue Cell Lysate lysate 1:2 Average % of 87 88 Expected Range ( %) 77-99 78-101 1:4 Average % of 89 94 Expected Range ( %) 78-103 82-104
    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
  • Williams, Jiang, Cochran, Dorsam, Graves: "Regulated expression of monocyte chemoattractant protein-1 in normal human osteoblastic cells." in: The American journal of physiology, Vol. 263, Issue 1 Pt 1, pp. C194-9, (1992) (PubMed).

    Beall, Mahajan, Kolattukudy: "Conversion of monocyte chemoattractant protein-1 into a neutrophil attractant by substitution of two amino acids." in: The Journal of biological chemistry, Vol. 267, Issue 5, pp. 3455-9, (1992) (PubMed).

  • 抗原 See all CCL2 ELISA试剂盒
    CCL2 (Chemokine (C-C Motif) Ligand 2 (CCL2))
    别名
    MCP-1 / CCL2 (CCL2 产品)
    别名
    GDCF-2 ELISA Kit, HC11 ELISA Kit, HSMCR30 ELISA Kit, MCAF ELISA Kit, MCP-1 ELISA Kit, MCP1 ELISA Kit, SCYA2 ELISA Kit, SMC-CF ELISA Kit, AI323594 ELISA Kit, JE ELISA Kit, Scya2 ELISA Kit, Sigje ELISA Kit, MCP-1A ELISA Kit, MCP1A ELISA Kit, C-C motif chemokine ligand 2 ELISA Kit, chemokine (C-C motif) ligand 2 ELISA Kit, C-C motif chemokine 2 ELISA Kit, CCL2 ELISA Kit, Ccl2 ELISA Kit, LOC101120093 ELISA Kit
    背景
    Monocyte chemoattractant protein-1 (MCP-1), also called monocyte chemotactic protein-1, belongs to the family of Chemokines. MCP-1 is expressed by monocytes, vascular endothelial cells, smooth muscle cells, glomerular mesangial cell, and osteoblastic cells. It has been shown to exhibit biological activities other than Chemotaxis. The activity of mouse MCP-1 has been shown to be mediated by the mouse CC chemokine receptor CCR2. The Mouse MCP-1 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse MCP-1 cell lysate and tissue lysate. This assay employs an antibody specific for mouse MCP-1 coated on a 96-well plate. Standards and samples are pipetted into the wells and MCP-1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse MCP-1 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 MCP-1 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
    20296
    UniProt
    P10148
    途径
    Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, ER-Nucleus Signaling, Unfolded Protein Response, The Global Phosphorylation Landscape of SARS-CoV-2 Infection
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