Human hypoglycosylated IgA1 (UGIGA1) ELISA kit experimental principle - Database & Sql Blog Articles

EL-C1600N100013-B
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Test probe P100-M3

Human hypoglycosylated IgA1 (UGIGA1) ELISA Kit

This kit is intended for research use only. It employs a double-antibody sandwich immunoassay to detect the level of human hypoglycosylated IgA1 in biological samples.

The procedure involves coating a microplate with a specific monoclonal antibody against hypoglycosylated IgA1. The sample, containing the target antigen, is added to the wells and allowed to bind to the coated antibody. An HRP-labeled secondary antibody is then introduced, forming an immune complex. After washing away unbound components, the substrate TMB is added, leading to a color change that correlates with the concentration of hypoglycosylated IgA1 in the sample. The reaction is stopped, and the absorbance at 450 nm is measured using a microplate reader. A standard curve is used to calculate the actual concentration of the analyte in the sample.

Kit Components:

  • 130x concentrated washing solution – 20ml × 1 bottle
  • Stop solution – 6ml × 1 bottle
  • Enzyme reagent – 6ml × 1 bottle
  • Standard (16μg/ml) – 0.5ml × 1 bottle
  • Enzyme-labeled plate – 12 wells × 8
  • Sample diluent – 6ml × 1 bottle
  • TMB reagent A – 6ml × 1 bottle
  • TMB reagent B – 6ml × 1 bottle
  • Instructions – 1 copy
  • Sealing film – 2 sheets
  • Sealed bag – 1

Sample Requirements:

It is recommended to process the specimens as soon as possible after collection. If not tested immediately, samples should be stored at -20°C, avoiding repeated freeze-thaw cycles. Avoid using samples containing NaN3, as it may inhibit the activity of horseradish peroxidase (HRP).

Procedure Summary:

  1. Dilute the standard according to the provided instructions.
  2. Add standards, controls, and samples to the microplate.

Calculation:

Plot the standard curve using OD values versus known concentrations. Use linear regression or direct comparison to determine the sample concentration, adjusting for any dilution factor applied during the experiment.

Precautions:

  • Allow the kit to reach room temperature before use.
  • Ensure accurate pipetting and avoid cross-contamination.
  • Always prepare a standard curve and run duplicates if necessary.
  • Store all waste materials as biohazardous.
  • Do not mix reagents from different batches.

Storage:

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

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