Human vascular endothelial cell adhesion factor (VCAM) ELISA kit instructions - Database & Sql Blog Articles

Brand AVX TPSE226M035R0125 Low impedance tantalum capacitor AVX 22

Electronic scale crystal oscillator 3.2*2.5mm 3225 16M (16.000MHZ) 12PF 10PPM 20PPM 30PPM

Test - uppercase JPG - memory 146K

Kaixin micro test

Human vascular endothelial cell adhesion factor (VCAM)

ELISA Kit Instructions

This kit is for research use only and not intended for human or animal consumption.

Experimental Principle

The Human Vascular Cell Adhesion Molecule (VCAM) ELISA Kit is designed to measure the levels of VCAM in biological samples using a double-antibody sandwich method. The process involves coating a microplate with a specific antibody against VCAM, then adding the sample and a labeled secondary antibody. A colorimetric reaction occurs through the enzyme horseradish peroxidase (HRP), allowing quantification of VCAM based on the intensity of the color developed at 450 nm.

Kit Composition

1. 30× Washing Solution – 20ml × 1 bottle 2. Stop Solution – 6ml × 1 bottle 3. Enzyme Standard Reagent – 6ml × 1 bottle 4. Standard (1200μg/L) – 0.5ml × 1 bottle 5. Enzyme-labeled Coating Plate – 12 wells × 8 strips 6. Standard Diluent – 1.5ml × 1 bottle 7. Sample Diluent – 6ml × 1 bottle 8. Instruction Manual – 1 copy 9. Reagent A – 6ml × 1 bottle 10. Reagent B – 6ml × 1 bottle 11. Sealing Film – 2 sheets 12. Ziplock Bag – 1

Sample Requirements

1. Samples should be processed as soon as possible after collection. If not tested immediately, store at -20°C and avoid repeated freezing and thawing. 2. Avoid using samples containing NaN3, as it may inhibit HRP activity.

Procedure

1. Standard Dilution: Prepare standards by diluting the original standard according to the provided chart. 2. Loading: Add 50μl of standard, 40μl of sample diluent, and 10μl of sample into each well. 3. Incubation: Seal the plate and incubate at 37°C for 30 minutes. 4. Washing: Wash the plate 5 times with diluted washing solution. 5. Add Enzyme: Add 50μl of enzyme reagent to each well except blank wells. 6. Incubation: Repeat the incubation step. 7. Washing: Perform washing again. 8. Color Development: Add 50μl of TMB substrate and develop for 10 minutes at 37°C. 9. Stop Reaction: Add 50μl of stop solution to terminate the reaction. 10. Measurement: Read OD values at 450 nm within 15 minutes.

Calculation

Plot a standard curve using OD values vs. concentration. Use linear regression or direct comparison to calculate sample concentrations. Multiply by the dilution factor for accurate results.

Precautions

1. Allow the kit to reach room temperature before use. Store unopened enzyme reagents in sealed bags. 2. If washing solution crystallizes, heat gently to dissolve. 3. Use accurate pipettes and control loading time. 4. Always run a standard curve and consider diluting high-concentration samples. 5. Use one sealing film per experiment. 6. Protect the substrate from light. 7. Follow instructions carefully and rely on microplate reader readings. 8. Treat all waste as biohazardous materials. 9. Do not mix components from different batches.

Storage and Shelf Life

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