Human M-CSF ELISA Kit

Protocol Booklet

Key features and details

Specification: 96 Test
Sensitivity: 1.41 pg/ml (50 μl)；15.89 pg/ml (10 μl)
Standard Curve Range: 78.13~5000 pg/ml
Standard Curve Gradient: 7 Points
Number of Incubations: 2
Sample Volume: 50 μl/10 μl
Assay type: Sandwich Elisa
Operation Duration: 120min

Brand:

CAT.NO. : AEH0126

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Size:

96T

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Product Details

Background

M-CSF, also known as CSF-1, is a four-alpha -helical-bundle cytokine that is the primary regulator of macrophage survival, proliferation and differentiation. M-CSF protein is also essential for the survival and proliferation of osteoclast progenitors. M-CSF also primes and enhances macrophage killing of tumor cells and microorganisms, regulates the release of cytokines and other inflammatory modulators from macrophages, and stimulates pinocytosis. M-CSF increases during pregnancy to support implantation and growth of the decidua and placenta. Sources of M-CSF include fibroblasts, activated macrophages, endometrial secretory epithelium, bone marrow stromal cells and activated endothelial cells . The M-CSF receptor (c-fms) transduces its pleotropic effects and mediates its endocytosis. M-CSF mRNAs of various sizes occur. Full length mouse M-CSF transcripts encode a 520 amino acid (aa) type I transmembrane (TM) protein with a 462 aa extracellular region, a 21 aa TM domain, and a 37 aa cytoplasmic tail that forms a 140 kDa covalent dimer. Differential processing produces two proteolytically cleaved, secreted dimers. One is an N- and O- glycosylated 86 kDa dimer, while the other is modified by both glycosylation and chondroitin-sulfate proteoglycan (PG) to generate a 200 kDa subunit. Although PG-modified M-CSF protein can circulate, it may be immobilized by attachment to type V collagen . Shorter transcripts encode M‑CSF that lacks cleavage and PG sites and produces an N-glycosylated 68 kDa TM dimer and a slowly produced 44 kDa secreted dimer. Although forms may vary in activity and half-life, all contain the N-terminal 150 aa portion that is necessary and sufficient for interaction with the M-CSF receptor. The first 229 aa of mature mouse M-CSF shares 87%, 83%, 82% and 81% aa identity with corresponding regions of rat, dog, cow and human M-CSF, respectively. Human M‑CSF is active in the mouse, but mouse M-CSF is reported to be species-specific.

Typical data

pg/ml	O.D.		Average	Corrected
0.00	0.0157	0.0157	0.0157
78.13	0.0691	0.0610	0.0651	0.0494
156.25	0.1317	0.1102	0.1210	0.1053
312.50	0.2432	0.2484	0.2458	0.2301
625.00	0.4781	0.4755	0.4768	0.4611
1250.00	0.9490	0.9950	0.9720	0.9563
2500.00	2.0360	1.7810	1.9085	1.8928
5000.00	3.3130	3.2000	3.2565	3.2408

Precision

	Intra-assay Precision			Inter-assay Precision
Sample Number	S1	S2	S3	S1	S2	S3
Sample Number	22	22	22	6	6	6
Average（pg/ml）	165.6	689.8	2146.1	110.7	548.3	1660.5
Standard deviation	9.8	42.2	119.7	5.2	37.6	83.2
Coefficient of variation（%）	5.9	6.1	5.6	4.7	6.9	5.0

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays)Three samples of known concentration were tested six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of human M-CSF into health human serum sample. The un-spiked serum was used as blank in this experiment.

The recovery ranged from 72% to 121% with an overall mean recovery of 105%.

Sample Values

Sample Matrix

Sample Evaluated

Range (pg/ml)

Detectable

Mean of Detectable (pg/ml)

Serum

684.79-1505.39

100

1024.43

Serum/Plasma - Thirty samples from apparently healthy volunteers were evaluated for the presence of M-CSF in this assay.No medical histories were available for the donors.

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