Human CD85d ELISA Kit

Protocol Booklet

Key features and details

Specification: 96 Test
Sensitivity 10.40 pg/ml (50 μl)：14.80 pg/ml (10 μl)
Standard Curve Range 312.5~20000 pg/ml
Standard Curve Gradient 7 Points/3 Folds
Number Of Incubations 2
Sample Volume 50 μl/10 μl
Assay Type Sandwiich Elisa
Operation Duration 120min

Brand:

CAT.NO. : AEH0046

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Size:

96T

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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
Sample Number	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%.

Sample Values

Sample Matrix

Sample Evaluated

Range (ng/ml)

Detectable

Mean of Detectable (ng/ml)

Serum

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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