Human Renal Aminease (RNLS/MAO-C) ELISA Kit Instructions for Use - Database & Sql Blog Articles

Human Renal Aminease (RNLS/MAO-C)

ELISA

Kit

Instructions for Use

This kit is intended for research use only and not for diagnostic or therapeutic purposes.

Experimental Principle

The Human Renal Aminease (RNLS/MAO-C) ELISA Kit is designed to measure the concentration of RNLS/MAO-C in biological samples using a double-antibody sandwich immunoassay. The microtiter plate is pre-coated with a specific antibody against human RNLS/MAO-C. After adding the sample, the target antigen binds to the immobilized antibody. A horseradish peroxidase (HRP)-labeled secondary antibody then binds to the captured antigen, forming an immune complex. Following incubation and washing steps, the TMB substrate is added, which changes color under the catalytic action of HRP. The intensity of the color is directly proportional to the amount of RNLS/MAO-C in the sample. The absorbance is measured at 450 nm using a microplate reader, and the concentration is calculated based on a standard curve.

Kit Composition

1. 130× Washing Solution – 20 ml × 1 bottle 2. Stop Solution – 6 ml × 1 bottle 3. Enzyme Standard Reagent – 6 ml × 1 bottle 4. Standard (640 ng/L) – 0.5 ml × 1 bottle 5. Enzyme-Labeled Coating Plate – 12 wells × 8 plates 6. Sample Diluent – 6 ml × 1 bottle 7. TMB Substrate A – 6 ml × 1 bottle 8. TMB Substrate B – 6 ml × 1 bottle 9. Standard Dilutions – 1.5 ml × 1 bottle 10. Instructions – 1 copy 11. Sealing Film – 2 sheets 12. Sealed Bag – 1

Sample Requirements

1. Samples should be processed as soon as possible after collection. If testing cannot be performed immediately, store samples at -20°C, avoiding repeated freeze-thaw cycles. 2. Avoid using samples containing NaN₃, as it may inhibit the activity of horseradish peroxidase (HRP).

Testing Procedure

1. Standard Dilution: Prepare a series of standards from the original stock solution according to the provided dilution chart. - 320 ng/L (No. 5) - 160 ng/L (No. 4) - 80 ng/L (No. 3) - 40 ng/L (No. 2) - 20 ng/L (No. 1) 2. Loading: Add 50 µL of each standard and 40 µL of sample diluent to the corresponding wells, followed by 10 µL of the sample. Ensure proper mixing. 3. Incubation: Seal the plate and incubate at 37°C for 30 minutes. 4. Washing: Wash the plate five times with diluted washing buffer (1:30). 5. Enzyme Addition: Add 50 µL of enzyme-labeled reagent to each well except blank controls. 6. Incubation: Repeat the incubation step at 37°C for 30 minutes. 7. Washing: Perform another five washes with the washing buffer. 8. Color Development: Add 50 µL of TMB A and 50 µL of TMB B to each well. Incubate at 37°C for 10 minutes. 9. Stop Reaction: Add 50 µL of stop solution to each well to halt the reaction. 10. Measurement: Read the absorbance at 450 nm within 15 minutes of stopping the reaction.

Data Analysis

Plot the standard curve using the OD values and concentrations. Calculate the sample concentration by interpolating the OD value from the standard curve. Multiply by the dilution factor to obtain the actual concentration.

Precautions

1. Allow the kit to reach room temperature before use. Store unopened enzyme reagents in a sealed bag. 2. If the washing solution crystallizes, warm it gently in a water bath before use. 3. Use a pipette for accurate measurements. Limit loading time to 5 minutes. 4. Always run a standard curve in duplicate. If the sample OD exceeds that of the highest standard, dilute the sample before testing. 5. Use a new sealing film for each test to prevent contamination. 6. Keep the substrate away from light. 7. Follow the instructions strictly and rely on microplate reader results. 8. Treat all waste materials as biohazardous. 9. Do not mix components from different batches.

Storage Conditions and Expiration

1. Store the kit at 2–8°C. 2. Shelf life is 6 months from the date of manufacture.