Human M-CSF ELISA Kit

Catalog No.: BEK1146

Size: 96T

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 10pg/ml

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

Application: For quantitative detection of M-CSF in tissue lysates or cell culture supernates, human serum and plasma should be validated by end users.

Introduction

Macrophage colony-stimulating factor, or M-CSF, is a secreted cytokine which influences hematopoietic stem cells to differentiate into macrophages or other related cell types. Ladner et al. (1987) showed that there are 2 forms of M-CSF, with 224 and 522 amino acids, resulting from alternative splicing. It is a hematopoietic growth factor that is involved in the proliferation, differentiation, and survival of monocytes, macrophages, and bone marrow progenitor cells. It released by osteoblasts (as a result of endocrine stimulation by parathyroid hormone) exerts paracrine effects on osteoclasts. M-CSF binds to receptors on osteoclasts inducing differentiation, and ultimately leading to increased plasma calcium levels—through the resorption (breakdown) of bone. More recently, it was discovered that CSF-1 and its receptor CSF1R are implicated in the mammary gland during normal development and neoplastic growth.

Principle of the Assay  

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

Kit components

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