Human GITRTNFRSF18 ELISA Kit

Protocol Booklet

Key features and details

Specification: 96 Test
Sensitivity: 0.07 pg/ml (50 μl)；1.20 pg/ml (10 μl)
Standard Curve Range: 5.49~4000 pg/ml
Standard Curve Gradient: 7 Points/3 Folds
Number of Incubations: 2
Sample Volume: 50 μl/10 μl
Assay type: Sandwich Elisa
Operation Duration: 120min

Brand:

CAT.NO. : AEH0095

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Size:

96T

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Product Details

Background

GITR (glucocorticoid-induced tumor necrosis factor receptor, also named AITR, activation-inducible TNF R family member), is a 228 amino acid (aa) type I transmembrane protein belonging to the TNF R family and has been designated TNFRSF18. The GITR cytoplasmic domain has striking homology with the cytoplasmic domain of 4-1BB and CD27. Human GITR shares 55% homology with murine GITR. GITR is expressed at low levels in peripheral blood T cells, bone marrow, thymus, spleen, and lymph nodes. In contrast to mouse GITR, expression of human GITR is not induced by treatment with dexamethasone, but is up-regulated by antigen stimulation or by treatment with anti-CD3 plus anti-CD28, or PMA plus ionomycin. Human GITR ligand was identified from human umbilical vein endothelial cells and is a 177 aa polypeptide belonging to the TNF superfamily (TNFSF18). Ligation of GITR can activate NF-kappa B through TRAF2, and protect T cells from TCR activation-induced cell death. It has been proposed that GITR ligand and GITR may modulate T lymphocyte functions.

Typical Data

pg/ml	O.D.		Average	Corrected
0.00	0.0145	0.0146	0.0146
5.49	0.0260	0.0256	0.0258	0.0113
16.46	0.0492	0.0497	0.0495	0.0349
49.38	0.1206	0.1200	0.1203	0.1058
148.15	0.3372	0.3175	0.3274	0.3128
444.44	0.9332	0.9130	0.9231	0.9086
1333.33	2.4945	2.4639	2.4792	2.4647
4000.00	4.1397	4.2936	4.2167	4.2021

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）	1410.0	535.3	111.0	1556.0	680.1	136.7
Standard deviation	55.5	10.1	2.4	102.5	32.1	9.1
Coefficient of variation（%）	3.9	1.9	2.2	6.6	4.7	6.7

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 GITR/TNFRSF18 into health serum sample. The un-spiked serum was used as blank in this experiment.

The recovery ranged from 84% to 111% with an overall mean recovery of 98%.

Sample Values

Sample Matrix

Sample Evaluated

Range (pg/ml)

Detectable

Mean of Detectable (pg/ml)

Serum

n.d.-1.19

0.47

Serum/Plasma - Thirty samples from apparently healthy volunteers were evaluated for the presence of human GITR/TNFRSF18 in this assay. No medical histories were available for the donors.

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