| Description |
The Human eosinophil cationic protein (ECP) ELISA Kit is engineered for accurate measurement of ECP levels from samples including serum, plasma, and tissue homogenates. It uses the Sandwich-ELISA mechanism in combination with the enzyme-substrate chromogenic reaction to measure the ECP content in the sample. ECP, also known as RNase-3, is a toxic granulocyte-derived protein releases by eosinophils upon its activation. ECP has marked toxicity for numerous helminth parasites, hemoflagellates, bacteria, single-stranded RNA viruses, and host tissues. Serum ECP can be used as a clinical tool for estimating eosinophil inflammatory activity in asthma and other allergic diseases. The levels of ECP are associated with disease severity and the extent of the achieved asthma control. In vitro, ECP can function as an antiviral agent and may be involved in host defense against the single-stranded RNA respiratory syncytial virus. The RNase activity of ECP is required for its antiviral and neurotoxic activities.
The kit has been validated against standards of sensitivity, specificity, precision, linearity, recovery, and lot-to-lot consistency. See more details through the product instructions.
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| Target Name |
eosinophil cationic protein,ECP |
| Alternative Names |
Cytotoxic ribonuclease ELISA Kit; ECP ELISA Kit; ECP_HUMAN ELISA Kit; Eosinophil cationic protein ELISA Kit; OTTHUMP00000164017 ELISA Kit; Ribonuclease 3 ELISA Kit; Ribonuclease; RNase A family; 3 ELISA Kit; RNase 3 ELISA Kit; RNASE3 ELISA Kit; RNS3 ELISA Kit |
| Abbreviation |
ECP |
| Species |
Homo sapiens (Human) |
| Sample Types |
serum, plasma, tissue homogenates |
| Detection Range |
1.56 ng/mL-100 ng/mL |
| Sensitivity |
0.39 ng/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Others |
| Assay Principle |
quantitative |
| Measurement |
Sandwich |
| 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 human ECP 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 % |
88-98 |
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| 1:2 |
Average % |
97 |
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| Range % |
92-103 |
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| 1:4 |
Average % |
100 |
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| Range % |
94-108 |
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| 1:8 |
Average % |
108 |
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| Range % |
103-112 |
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| Recovery |
| The recovery of human ECP 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) |
90 |
87-95 |
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| EDTA plasma (n=4) |
92 |
89-95 |
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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. |
| ng/ml |
OD1 |
OD2 |
Average |
Corrected |
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| 100 |
2.500 |
2.493 |
2.497 |
2.392 |
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| 50 |
2.003 |
1.998 |
2.001 |
1.896 |
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| 25 |
1.275 |
1.266 |
1.271 |
1.166 |
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| 12.5 |
0.587 |
0.573 |
0.580 |
0.475 |
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| 6.25 |
0.369 |
0.357 |
0.363 |
0.258 |
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| 3.12 |
0.221 |
0.208 |
0.215 |
0.110 |
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| 1.56 |
0.176 |
0.169 |
0.173 |
0.068 |
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| 0 |
0.108 |
0.102 |
0.105 |
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| Materials provided |
- A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human ECP antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard --- Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled ECP antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the high concentration Biotin-antibody to an appropriate working solution.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the high concentration HRP-avidin solution to an appropriate solution.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) --- Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) --- Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- 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 540 nm or 570 nm.
- An incubator that can provide stable incubation conditions up to 37°C±5°C.
- 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 |
