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

Fish Total Lipase (TLP) ELISA Kit Instructions This kit is intended for research purposes only and is designed to measure the total lipase (TLP) levels in fish serum, plasma, and related liquid samples. The detection range of this assay is from 12 pg/ml to 450 pg/ml. **Principle of the Assay** The Fish Total Lipase (TLP) ELISA Kit utilizes a double-antibody sandwich method. A microtiter plate is coated with purified TLP antibodies, forming a solid-phase antibody. After adding the sample, TLP binds to the immobilized antibody. HRP-labeled TLP antibody is then added, forming an immune complex. Following washing, TMB substrate is introduced, and the reaction is stopped with an acidic solution. The color change from blue to yellow is proportional to the TLP concentration in the sample. The absorbance is measured at 450 nm using a microplate reader, and the TLP concentration is determined by comparing the OD values to a standard curve. **Kit Components** - 30× Washing Solution: 20 ml × 1 bottle - Stop Solution: 6 ml × 1 bottle - Enzyme Standard Reagent: 6 ml × 1 bottle - Standard Product (800 pg/ml): 0.5 ml × 1 bottle - Enzyme-Labeled Coating Plate: 12 wells × 8 strips - Sample Diluent: 6 ml × 1 bottle - Standard Diluent: 1.5 ml × 1 bottle - TMB Developer A: 6 ml × 1 bottle - TMB Developer B: 6 ml × 1 bottle - Instruction Manual: 1 copy - Seal Film: 2 sheets - Sealing Bag: 1 piece **Sample Requirements** Samples should be processed as soon as possible after collection. If not tested immediately, store at -20°C and avoid repeated freezing and thawing. Samples containing NaN3 are not suitable for testing due to potential inhibition of HRP activity. **Procedure** 1. **Standard Dilution**: Prepare standards according to the provided dilution chart. 2. **Sample Addition**: Add 50 μl of standard or sample diluent (40 μl) and 10 μl of sample to each well. 3. **Incubation**: Seal the plate and incubate at 37°C for 30 minutes. 4. **Washing**: Wash the plate five times using diluted washing solution. 5. **Enzyme Addition**: Add 50 μl of enzyme-labeled reagent to all wells except blank. 6. **Second Incubation**: Repeat the incubation step. 7. **Color Development**: Add 50 μl of TMB A and B, mix gently, and incubate at 37°C for 10 minutes. 8. **Stop Reaction**: Add 50 μl of stop solution to each well. 9. **Measurement**: Read OD values at 450 nm within 15 minutes. **Data Analysis** Plot the standard curve using standard concentrations and corresponding OD values. Determine the sample concentration based on the standard curve and multiply by the dilution factor. **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 test. - Keep substrates away from light. - Always follow the manual carefully. - Treat all samples and waste as biohazardous materials. - Do not mix components from different batches. **Storage and Shelf Life** - Store the kit at 2–8°C. - Shelf life: 6 months from the date of manufacture.

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