Human IFN-gamma R1 ELISA Kit

Protocol Booklet

Key features and details

Specification: 96 Test
Sensitivity: 2.28 pg/ml (50 μl)；11.4 pg/ml (10 μl)
Standard Curve Range: 13.72~10000 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. : AEH0104

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Size:

96T

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Product Details

Background

The high-affinity IFN-gamma receptor complex is made up of two type I membrane proteins, IFN-gamma R1 (IFN-gamma R alpha ) and IFN-gamma R2 (IFN-gamma R beta ). Both proteins are members of the type II cytokine receptor family and share approximately 52% amino acid sequence identity. IFN-gamma R1 is the ligand-binding subunit that is necessary and sufficient for IFN-gamma binding and receptor internalization. IFN-gamma R2 is required for IFN-gamma signaling but does not bind IFN-gamma by itself. Human IFN-gamma R1 cDNA encodes a 499 amino acid (aa) residue protein with a 17 aa signal peptide, a 228 aa extracellular domain, a 23 aa transmembrane domain, and a 221 aa intracellular domain. Human and mouse IFN-gamma R1 share 52% amino acid sequence identity and bind IFN-gamma in a species-specific manner. IFN-gamma R1 is constitutively expressed in most cell types. Soluble IFN-gamma R1 that binds IFN-gamma has been detected in biological fluids.

Typical Data

pg/ml	O.D.		Average	Corrected
0.00	0.0285	0.0286	0.0286
13.72	0.0405	0.0381	0.0393	0.0108
41.15	0.0631	0.0567	0.0599	0.0314
123.46	0.1253	0.1176	0.1215	0.0929
370.37	0.3195	0.2907	0.3051	0.2766
1111.11	0.8572	0.7749	0.8161	0.7875
3333.33	2.1650	2.0410	2.1030	2.0745
10000.00	4.0214	3.8750	3.9482	3.9197

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）	216.4	1044.3	3466.9	188.9	972.7	3239.0
Standard deviation	9.7	50.3	148.6	15.9	64.1	77.7
Coefficient of variation（%）	4.5	4.8	4.3	4.4	6.6	2.4

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 IFN gamma R1 into health human serum sample. The un-spiked serum was used as blank in this experiment. The recovery ranged from 93% to 120% with an overall mean recovery of 103%.

Sample Values

Sample Matrix

Sample Evaluated

Range (pg/ml)

Detectable

Mean of Detectable (pg/ml)

Serum

284.50-993.11

100

576.85

Serum/Plasma - Thirty samples from apparently healthy volunteers were evaluated for the presence of human IFN gamma R1 in this assay. No medical histories were available for the donors.

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