Human GM-CSF ELISA Kit

Catalog No.: BEK1071

Size: 96T

Range: 7.8 pg/ml - 500 pg/ml

Sensitivity < 2 pg/ml

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

Application: For quantitative detection of GM-CSF in human serum, plasma, body fluids, tissue lysates or cell culture supernates.

Introduction

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a protein secreted by macrophages, T cells, mast cells, NK cells, endothelial cells and fibroblasts. It is essential for the in vitro proliferation and differentiation of precursor cells into mature granulocytes and macrophages. GM-CSF stimulates stem cells to produce granulocytes (neutrophils, eosinophils, and basophils) and monocytes. GM-CSF is used as a medication to stimulate the production of white blood cells following chemotherapy. Recently, it has been evaluated in clinical trials for its potential as a vaccine adjuvant in HIV-infected patients. The preliminary results have been promising but GM-CSF is not presently FDA-approved for this purpose.

Principle of the Assay 

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-GM-CSF polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-GM-CSF 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 GM-CSF amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of GM-CSF can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human GM-CSF antibody
2. Lyophilized Human GM-CSF standards: 2 tubes (10 ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human GM-CSF 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