Human CD85d ELISA Kit
Protocol Booklet 
Key features and details
- Species:
- Size:
- Sample Volume:
- Standard Curve Range:
- Assay type:
- Sensitivity:
- Operation Duration:
- Assay type:
-
Brand:
CAT.NO. : AEH0046
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Product Details
Background
The immunoglobulin-like transcript (ILT) comprise a family of activating and inhibitory type immunoreceptors whose genes are located in the same locus that encodes killer cell Ig-like receptors (KIR). ILT4, also known as LIR-2 and LILRB2, is a type I transmembrane protein expressed primarily on monocytes and dendritic cells (DC). Human ILT4 is produced as a 598 amino acid (aa) precursor including a 21 aa signal sequence, a 440 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 116 aa cytoplasmic domain. The ECD contains four Ig-like domains, and the cytoplasmic domain contains three immunoreceptor tyrosine-based inhibitory motifs (ITIM). The ECD of human ILT4 shares 76% aa identity with chimpanzee ILT4 and 74%, 81%, 33%, 52%, 77%, 61%, and 64 % aa identity with human ILT1, 2, 3, 5, 6, 7, and 8, respectively. ILT4 binds to classical MHC I proteins as well as the non-classical HLA-G1 and HLA-F molecules. It competes with CD8 alpha for MHC I binding but does not compete with KIR2DL1. Ligation of ILT4 induces Tyr phosphorylation within its cytoplasmic ITIMs, a requirement for association with SHP-1. Activation of ILT4 inhibits signaling through Fc gamma RI and Fc epsilon RI and causes DC to become tolerogenic by downregulation of costimulatory molecules. ILT4 mediates tolerogenic DC-induced CD4+ T cell energy in vitro and in vivo.
Typical data
|
pg/ml |
O.D. |
Average |
Corrected |
|
|
0.00 |
0.0345 |
0.0329 |
0.0337 |
|
|
312.50 |
0.0936 |
0.0922 |
0.0929 |
0.0592 |
|
625.00 |
0.1418 |
0.1472 |
0.1445 |
0.1108 |
|
1250.00 |
0.2482 |
0.2518 |
0.2500 |
0.2163 |
|
2500.00 |
0.4908 |
0.4994 |
0.4951 |
0.4614 |
|
5000.00 |
0.9079 |
0.9281 |
0.9180 |
0.8843 |
|
10000.00 |
1.8010 |
1.8280 |
1.8145 |
1.7808 |
|
20000.00 |
3.2730 |
3.2200 |
3.2465 |
3.2128 |
Precision
|
Intra-assay Precision |
Inter-assay Precision |
|||||
|
Sample Number |
S1 |
S2 |
S3 |
S1 |
S2 |
S3 |
|
22 |
22 |
22 |
6 |
6 |
6 |
|
|
Average（pg/ml） |
278.4 |
1676.4 |
5340.2 |
335.2 |
1676.4 |
5347.2 |
|
Standard deviation |
18.3 |
50.3 |
193.0 |
17.5 |
80.8 |
160.4 |
|
Coefficient of variation（%） |
6.6 |
3.0 |
3.6 |
5.2 |
4.8 |
3.0 |
- 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 CD85d into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 80% to 127% with an overall mean recovery of 112%.
The recovery ranged from 80% to 127% with an overall mean recovery of 112%.
Sample values
|
Sample Matrix |
Sample Evaluated |
Range (ng/ml) |
Detectable % |
Mean of Detectable (ng/ml) |
|
Serum |
30 |
2.46-9.25 |
100 |
5.35 |
Serum/Plasma - Thirty samples from apparently healthy volunteers were evaluated for the presence of human CD85d in this assay. No medical histories were available for the donors.
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