Mouse LIF enzyme-linked immunoassay kit

CAT: EM0038 Datasheet
Specification 96 Test
Sensitivity 2.77 pg/ml (10 μl)
Standard Curve Range 6.86~5000 pg/ml
Standard Curve Gradient 7 Points/3 Folds
Number of Incubations 2
Sample Volume 10 μl
Type Fully Ready-to-Use
Operation Duration 120min
pg/ml O.D. Average Corrected
0.00 0.0337 0.0309 0.0323
6.86 0.0458 0.0440 0.0449 0.0126
20.58 0.0698 0.0664 0.0681 0.0358
61.73 0.1404 0.1345 0.1375 0.1052
185.19 0.3507 0.3391 0.3449 0.3126
555.56 0.9113 0.9009 0.9061 0.8738
1666.67 2.1080 2.0780 2.0930 2.0607
5000.00 3.6630 3.7060 3.6845 3.6522

Precision

Intra-assay Precision Inter-assay Precision
Sample Number S1 S2 S3 S1 S2 S3
22 22 22 6 6 6
Average(pg/ml) 102.3 501.8 1489.4 100.7 491.3 1511.1
Standard Deviation 3.2 20.1 62.0 3.6 15.2 44.2
Coefficient of Variation(%) 3.1 4.0 4.2 3.5 3.1 2.9

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 mouse LIF into health mouse serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 94% to 105% with an overall mean recovery of 100%.

Sample Values

Sample Matrix Sample Evaluated Range (pg/ml) Detectable (%) Mean of Detectable (pg/ml)
Serum 30 133.17-389.78 100 261.5

Serum/Plasma – Thirty samples from apparently healthy mice were evaluated for the presence of CRP in this assay. No medical histories were available for the donors. n.d. = non-detectable. Samples measured below the sensitivity are considered to be non-detectable.

Background: LIF

Recombinant mouse LIF (leukemia inhibitory factor) is commonly used in cell culture to maintain the pluripotency of stem cells. LIF is a widely expressed pleiotropic member of the IL-6 family of cytokines. Mature mouse LIF is expressed as a highly and variably glycosylated 32-62 kDa monomer that shares 78%, 91%, 80%, 76%, and 78% aa sequence identity with human, rat, canine, bovine, and porcine LIF, respectively. LIF functions through a heterodimeric receptor complex containing a ligand-binding subunit, LIF R alpha /CD118, and a signal transducing subunit, gp130. gp130 also serves as a subunit of the receptor complexes for Oncostatin M, Cardiotrophin-1, CNTF, IL-6, IL-11, and IL-27. A soluble form of mouse LIF R alpha can be generated by alternative splicing. Depending on the cells and their context, LIF either opposes or favors differentiation. LIF produced by the uterine endometrium supports successful implantation of the embryo, promotes proliferation and maintenance of pluripotency in embryonic stem cells, and favors proliferation of progenitor cell types such as hematopoietic stem cells. However, excess LIF blocks differentiation of embryoid bodies, indicating the importance of LIF regulation. LIF is produced by activated CD4+ T cells and is required by the thymic epithelium to support T cell maturation. Its expression is upregulated by neuronal injury, and it promotes motor neuron survival and oligodendrocyte myelination. It is produced by the adrenal cortex and likely enhances adrenal production of cortisol and aldosterone. LIF can also function as an autocrine growth factor in some pancreatic cancers, but it induces differentiation in the myeloid leukemic cell line M1. Tumor cell-derived LIF can also induce formation of immunosuppressive tumor-associated macrophages. LIF promotes endometrial remodeling and differentiation of adipocytes and cardiac smooth muscle cells. It promotes regulatory T cell and inhibits Th17 cell differentiation, thus down-regulating inflammation and contributing to immune tolerance during pregnancy and in the nervous system.

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