Human CD137/4-1BB/TNFRSF9 ELISA Kit

Protocol Booklet

Key features and details

Specification: 96 Test
Sensitivity: 0.47 pg/ml (50 μl)；5.07 pg/ml (10 μl)
Standard Curve Range: 15.63~1000 pg/ml
Standard Curve Gradient: 7 Points
Number of Incubations: 2
Sample Volume: 50 μl/10 μl
Assay type: Sandwich Elisa
Operation Duration: 120min

Brand:

CAT.NO. : AEH0124

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Size:

96T

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Product Details

Background

4-1BB is an inducible T cell surface protein belonging to the TNF receptor superfamily. It is alternatively known as TNFRSF9, CD137, and ILA. The 255 amino acid human 4-1BB is a type I transmembrane protein having in its extracellular domain four of the cysteine-rich motifs that are characteristic of the TNF receptor superfamily. The 30 kDa glycoprotein exists both as a monomer and as a dimer on T cells. The human and mouse proteins share 60% amino acid identity. 4-1BB is absent from naive T cells, but it is upregulated and continually expressed following T cell activation. The natural ligand, 4-1BBL, is a member of the TNF superfamily and is expressed on activated antigen presenting cells including dendritic cells, macrophages, and B cells. Cross-linking of 4-1BB by 4-1BBL or by agonistic antibodies transmits a potent co-stimulatory signal that enhances the effect of other activating signals such as PHA or anti-CD3 antibodies. 4-1BB signals through the TFAF2‑NIK pathway resulting in activation of NF-kappa B and ultimately promoting proliferation and survival of T cells.

Typical data

pg/ml	O.D.		Average	Corrected
0.00	0.0659	0.0698	0.0679
15.63	0.1978	0.2148	0.2063	0.1385
31.25	0.3346	0.3512	0.3429	0.2751
62.50	0.5916	0.6533	0.6225	0.5546
125.00	1.1130	1.1470	1.1300	1.0622
250.00	1.8960	1.9670	1.9315	1.8637
500.00	3.2850	3.4530	3.3690	3.3012
1000.00	4.3356	4.3766	4.3561	4.2883

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）	25.6	117.0	352.9	22.8	110.2	319.1
Standard deviation	1.4	3.5	10.2	1.1	2.3	12.4
Coefficient of variation（%）	5.3	3.0	2.9	4.7	2.1	3.9

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 human CD137/4-1BB/TNFRSF9 into health human serum sample. The un-spiked serum was used as blank in this experiment.

The recovery ranged from 86% to 116% with an overall mean recovery of 103%.

Sample Values

Sample Matrix

Sample Evaluated

Range (Pg/ml)

Detectable

Mean of Detectable (Pg/ml)

Serum

20.14-49.28

100

32.58

Serum/Plasma - Thirty samples from apparently healthy volunteers were evaluated for the presence of CD137/4-1BB/TNFRSF9 in this assay.No medical histories were available for the donors.

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