Fish immunoglobulin M (IgM) ELISA Kit
Product Details
| Description |
The fish immunoglobulin M (IgM) ELISA kit is suitable for the quantitative measurement of the IgM levels in fish serum, plasma, or tissue homogenates. It provides a detection range of 1.25 μg/mL-50 μg/mL. It also has high sensitivity, excellent specificity, high precision, good linearity, and high recovery. This assay employs the competitive inhibition enzyme immunoassay technique using an anti-fish IgM antibody and an HRP-conjugated IgM. In this assay, the microplate has been pre-coated with a goat-anti-rabbit antibody. Samples or standards are pipetted into wells with the anti-fish IgM antibody and HRP-conjugated IgM. IgM in the samples or the standards and HRP-conjugated IgM competes with binding to the anti-fish IgM antibody. The solution appears blue after the addition of the TMB substrate. The color reaction is terminated after adding the stop solution. The color intensity is inverse to the amount of IgM in the sample. IgM is the initial antibody produced by B cells in response t... Read more |
| Target Name |
immunoglobulin M |
| Abbreviation |
IgM |
| Species |
Fish |
| Sample Types |
serum, plasma, tissue homogenates |
| Detection Range |
1.25 μg/mL-50 μg/mL |
| Sensitivity |
1.25 μg/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Immunology |
| Assay Principle |
quantitative |
| Measurement |
Competitive |
| Precision |
| Intra-assay Precision (Precision within an assay): CV% |
| Three samples of known concentration were tested twenty times on one plate to assess. |
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| Inter-assay Precision (Precision between assays): CV% |
| Three samples of known concentration were tested in twenty assays to assess. |
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| Linearity |
| To assess the linearity of the assay, samples were spiked with high concentrations of fish IgM in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
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Sample |
Serum(n=4) |
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| 1:1 |
Average % |
93 |
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| Range % |
86-101 |
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| 1:2 |
Average % |
97 |
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| Range % |
90-104 |
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| 1:4 |
Average % |
95 |
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| Range % |
84-105 |
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| 1:8 |
Average % |
95 |
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| Range % |
91-99 |
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| Recovery |
| The recovery of fish IgM spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. |
| Sample Type |
Average % Recovery |
Range |
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| Serum (n=5) |
97 |
92-105 |
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| EDTA plasma (n=4) |
95 |
91-102 |
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| Typical Data |
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
| μg/ml |
OD1 |
OD2 |
Average |
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| 50 |
0.122 |
0.124 |
0.123 |
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| 25 |
0.213 |
0.199 |
0.206 |
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| 11.5 |
0.401 |
0.427 |
0.414 |
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| 3.75 |
0.745 |
0.761 |
0.753 |
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| 1.25 |
1.375 |
1.402 |
1.389 |
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| 0 |
2.541 |
2.532 |
2.537 |
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| Materials provided |
- A 96-well Assay plate --The 96-well plate has been pre-coated with goat-anti-rabbit antibody.
- Standard (6 x 0.5 ml) --Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
- HRP-conjugated IgM (1 x 6 ml) --Bind to the IgM antibody, and HRP catalyzes the TMB to elicit a chromogenic reaction.
- IgM Antibody (1 x 6 ml) --Interact with the IgM.
- Sample Diluent (2 x 20 ml) --Reconstitute the standard and dilute the sample to an appropriate concentration.
- Wash Buffer (25x concentrate) (1 x 20 ml) --Wash away unbound or free substances.
- TMB Substrate (1x 10 ml) --Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- Stop Solution (1 x 10ml) --Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells)
- An Instruction manual
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| Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 600 nm - 630 nm.
- An incubator that can provide stable incubation conditions up to 37°C±5°
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Timer
- Test tubes for dilution
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| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
