Fish total lipase (TLP) Elisa kit instructions - Database & Sql Blog Articles

**Fish Total Lipase (TLP) ELISA Kit Instructions** This kit is intended for research use only. It allows the quantitative detection of total lipase (TLP) in fish serum, plasma, and related liquid samples. The detection range is from 12 pg/ml to 450 pg/ml. **Experimental Principle** The Fish Total Lipase (TLP) ELISA Kit utilizes a double-antibody sandwich method. A microtiter plate is coated with purified TLP antibodies to create a solid-phase antibody. The sample containing TLP is added to the wells, followed by HRP-labeled TLP antibodies. This forms an antibody-antigen-enzyme-labeled antibody complex. After washing, the substrate TMB is added, which turns blue under the action of HRP and then changes to yellow when acid is introduced. The color intensity is directly proportional to the TLP concentration in the sample. The absorbance at 450 nm is measured using a microplate reader, and the TLP concentration is determined based on a standard curve. **Kit Composition** - 1 × 30× concentrated washing solution (20 ml/bottle) - 1 × Stop solution (6 ml/bottle) - 1 × Enzyme standard reagent (6 ml/bottle) - 1 × Standard product (800 pg/ml, 0.5 ml/bottle) - 1 × Enzyme-labeled coating plate (12 wells × 8 strips) - 1 × Sample dilution (6 ml/bottle) - 1 × Standard dilution (1.5 ml/bottle) - 1 × Instruction manual - 1 × Developer A (6 ml/bottle) - 1 × Developer B (6 ml/bottle) - 1 × Sealing film (2 sheets) - 1 × Sealed bag **Sample Requirements** Samples should be processed as soon as possible after collection, following relevant protocols. If not tested immediately, they can be stored at -20°C, but repeated freeze-thaw cycles should be avoided. Samples containing NaN3 are not suitable for testing, as it may inhibit horseradish peroxidase (HRP) activity. **Procedure** 1. **Standard Dilution**: Prepare standards according to the provided dilution chart. 2. **Sample Addition**: Add 50 μl of standard, 40 μl of sample diluent, and 10 μl of sample into designated wells. 3. **Incubation**: Seal the plate and incubate at 37°C for 30 minutes. 4. **Washing**: Use 30× diluted washing solution and wash the plate 5 times. 5. **Enzyme Addition**: Add 50 μl of enzyme-labeled reagent to each well except blank. 6. **Second Incubation**: Repeat incubation at 37°C for 30 minutes. 7. **Color Development**: Add 50 μl of TMB A and 50 μl of TMB B, incubate at 37°C for 10 minutes. 8. **Stop Reaction**: Add 50 μl of stop solution to terminate the reaction. 9. **Measurement**: Read OD values at 450 nm within 15 minutes. **Data Analysis** Plot the standard curve using OD values vs. concentrations. Calculate the sample concentration using linear regression or direct comparison. Multiply by the dilution factor to obtain the actual value. **Notes** - Allow the kit to equilibrate at room temperature before use. - Store unopened enzyme strips in a sealed bag. - Avoid cross-contamination by using a new sealing film for each experiment. - Keep substrates away from light. - Always follow the manual strictly and use a microplate reader for accurate results. - Dispose of all samples, waste, and reagents as biohazardous materials. - Do not mix components from different batches. **Storage Conditions** Store the kit at 2–8°C. The shelf life is 6 months from the date of manufacture.

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