Improve Research Reproducibility A Bio-protocol resource
Peer-reviewed
tooltip

Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Total Murine Immunoglobulin

Zheng Liu Zheng Liu

Published: Feb 5, 2011 DOI: 10.21769/BioProtoc.33 Views: 15185

download pdf Download PDF
ask Ask a question
Favorite
Cited by

Abstract

ELISA is an easy and relatively sensitive way to measure protein concentration. This protocol describes how to measure serum levels of murine total immunoglobulin of various isotypes. This protocol was developed or modified in Dr. Anne Davidson’s lab at Feinstein Institute for Medical Research.

Keywords: ELISA search Mouse search Immunoglobulin search Serum search

Materials and Reagents

  1. Coating antibody: goat anti-mouse IgX* unlabeled (Southern Biotech)
    Note: IgX*: the isotype of the Ig you wish to detect.
  2. Murine immunoglobulin of the desired isotype (Sigma- Aldrich, catalog number: M5284 )
  3. Horseradish peroxidase (HRP) conjugated goat anti-mouse isotype specific antibodies (Southern Biotech)
  4. 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) Peroxidase Substrate Solution A (Kirkegaard & Perry Laboratories, catalog number: 50-64-00 )
  5. ABTS Peroxidase Substrate Solution B (Kirkegaard & Perry Laboratories, catalog number: 50-65-00 )
  6. ABTS Peroxidase Stop Solution (Kirkegaard & Perry Laboratories, catalog number: 50-85-01 )
  7. 10x PBS-Tween 20 (see Recipes)
  8. Blocking solution (see Recipes)

Equipment

  1. Clear 96-well Microtest polystyrene assay plate (BD Biosciences)

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Immunology > Antibody analysis > Antibody detection

Biochemistry > Protein > Immunodetection

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A