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IgM ELISA 试剂盒

适用: 猴 Colorimetric Sandwich ELISA Plasma, Serum
产品编号 ABIN956142
发货至: 中国
  • 抗原 See all IgM ELISA试剂盒
    IgM
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    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    应用范围
    ELISA
    样品类型
    Plasma, Serum
    Analytical Method
    Quantitative
    特异性
    Cross-reactivity with immunoglobulins from other species has not been investigated.
    产品特性
    The monkey IgM ELISA kit is designed for measurement of IgM in old world monkey serum or plasma. The assay uses goat anti-monkey IgM for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated goat anti-monkey IgM antibodies for detection. Both capture and detection antibodies were cross-absorbed on monkey IgG and IgA agarose columns, thereby ensuring specificity for IgM.Test samples are diluted and incubated in the microtiter wells for 45 minutes alongside prepared monkey IgM calibrators. The microtiter wells are subsequently washed and HRP conjugate is added and incubated for 45 minutes. IgM molecules are thus sandwiched between the immobilization and detection antibodies. The wells are then washed to remove unbound HRP-labeled antibodies and TMB Reagent is added and incubated for 20 minutes at room temperature. This results in the development of a blue color. Color development is stopped by the addition of Stop Solution, changing the color to yellow, and optical density is measured spectrophotometrically at 450 nm. The concentration of IgM is proportional to the optical density of the test sample and is derived from a calibration curve.
    组件
    Anti-monkey IgM -coated microtiter wells, 96 wells
    Monkey IgM Calibrator (lyophilized)
    Diluent (10X), 50 mL
    HRP Conjugate, 11 mL
    Wash Solution (20X), 50 mL
    TMB Reagent (One-step), 11 mL
    Stop Solution (1N HCl), 11 mL.
    试剂未包括
    Precision pipettes and tips
    Distilled or de-ionized water
    Polypropylene or glass tubes
    Vortex mixer
    Absorbent paper or paper towels
    Micro-Plate incubator/shaker mixing speed of ~150 rpm
    A microtiter plate reader capable of measuring absorbance at 450 nm, with a bandwidth of 10 nm or less and an OD range of 0-4 OD
    Graph paper (PC graphing software is optional)
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    Discover our best selling IgM ELISA Kit
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    Discover our top product IgM ELISA Kit
  • 板类型
    Pre-coated
    样品制备

    General Note: IgM is typically present in monkey serum or plasma at concentrations of ~0.3 mg/mL. In order to obtain values within range of the calibration curve, we suggest that samples initially be diluted 10,000 fold using the following procedure for each sample to be tested:
    1. Dispense 247.5 µL of 1x diluent into two tubes.
    2. Pipette and mix 2.5 µL of the serum/plasma sample with 247.5 µL of 1x diluent in the first tube. This provides a 100 fold diluted sample.
    3. Mix 2.5 µL of the 100 fold diluted sample with the 247.5 µL of diluent in the second tube. This provides a 10,000 fold dilution of the sample.
    4. Repeat this procedure for each sample to be tested Tissue extracts and body fluids other than serum or plasma will likely have lower IgM levels than those found in serum. Optimal dilutions of such samples should be determined empirically.

    实验流程
    1. Secure the desired number of coated wells in the holder.
      2. Dispense 100 µL of calibrators and diluted samples into the wells (we recommend that samples be tested in duplicate).
      3. Incubate on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 45 minutes.
      4. Aspirate the contents of the microtiter wells and wash the wells 5 times with 1x wash solution using a plate washer (400 µL/well). The entire wash procedure should be performed as quickly as possible.
      5. Strike the wells sharply onto absorbent paper or paper towels to remove all residual wash buffer.
      6. Add 100 µL of enzyme conjugate reagent into each well.
      7. Incubate on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 45 minutes.
      8. Wash as detailed in 4 to 5 above.
      9. Dispense 100 µL of TMB Reagent into each well.
      10. Gently mix on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 20 minutes.
      11. Stop the reaction by adding 100 µL of Stop Solution to each well.
      12. Gently mix. It is important to make sure that all the blue color changes to yellow.
      13. Read the optical density at 450 nm with a microtiter plate reader within 5 minutes.
    结果分析
    1. Calculate the mean absorbance values (A450) for each set of reference calibrators and samples.
      2. Construct a calibration curve by plotting the mean absorbance obtained from each reference calibrator against its concentration in ng/mL on linear graph paper, with absorbance values on the vertical or Y-axis and concentration on the horizontal or X-axis.
      3. Using the mean absorbance value for each sample, determine the corresponding concentration of IgM in ng/mL from the calibration curve.
      4. Multiply the derived concentrations by the dilution factor to determine the actual concentration of IgM in the sample.
      5. PC graphing software may be used for the above steps.
      6. If the OD450 values of the sample fall outside the calibration curve, samples should be diluted appropriately and re-tested.
    限制
    仅限研究用
  • 储存条件
    4 °C
    储存方法
    Store kit at 4°C. Keep the microtiter plate in a sealed bag with desiccant to minimize exposure to damp air. The expiration date of the kit is indicated on the box label.
    有效期
    The expiry date is stated on the label.
  • 抗原 See all IgM ELISA试剂盒
    IgM
    Abstract
    IgM 产品
    别名
    IgM constant region ELISA Kit, IGM ELISA Kit
    物质类
    Antibody
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