Mouse CD30L ELISA Kit

Catalog No.: BEK1025

Size: 96T

Range: 15.6 pg/ml-1000 pg/ml

Sensitivity < 6 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of CD30L in mouse serum, body fluids, tissue lysates or cell culture supernatants.

Introduction

CD30 ligand (CD30L), also known as CD153, is a cytokine that induces proliferation of T cells. The CD30L gene contains 4 exons and spans more than 17.1 kb. It has the characteristics of a type II membrane protein, with no apparent signal peptide and a transmembrane domain followed by a C-terminal extracellular domain. CD30L is expressed on the surface of B cells and found that this expression is upregulated upon CD154 (CD40L), IL4, and B-cell receptor engagement. Smith et al. (1993) found that recombinant mouse CD30L enhanced the proliferation of CD3-activated T cells, but induced differential responses, including cell death, in several CD30-positive lymphoma-derived cell lines.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-CD30L polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-CD30L polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the CD30L amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of CD30L can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Mouse CD30L antibody
2. Lyophilized Mouse CD30L standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Mouse CD30L antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer(25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L