Human OX40/TNFRSF4 ARExSet® antibody pair for ELISA

Protocol Booklet

Human OX40/TNFRSF4 ARExSet® antibody pair for ELISA

Key features and details

Species: Human
Size: 5 plates,15 plates,50 plates
Sample Volume: 50 μl
Standard Curve Range: 46.88 pg/ml -3000 pg/ml
Assay type: Not Ready-to-Use
Sensitivity: 1.08 pg/ml(50 μl)
Operation Duration: 120min
Assay type: Sandwich Elisa

Brand:

CAT.NO. : ASEH0120

US$ Please choose

Size:

5 plates 15 plates 50 plates

Trail, Bulk size or Custom requests Please contact us

Product Details

Background

OX40 (CD134; TNFRSF4) is a T cell costimulatory molecule of the TNF receptor superfamily that coordinates with other membrane-bound costimulators such as CD28, CD40, CD30, CD27 and 4-1BB. OX40 is expressed on naïve CD4+ T cells only after engagement of the TCR by antigen presenting cells (APC; dendritic and B cells), and costimulation by CD40/CD40 ligand and CD28/B7. It is maximal at 2‑5 days post activation, or 4 hours post reactivation of memory T cells. Human OX40 is a 48 kDa type I transmembrane glycoprotein with a 28 amino acid (aa) signal sequence, a 185 aa extracellular domain (ECD) that has four TNFR-Cys repeats and an O-glycosylated hinge region, a 20 aa transmembrane segment, and a 41 aa cytoplasmic domain. The ECD of human OX40 shows 71%, 68%, 67%, 64% and 64% aa identity with feline, canine, rabbit, mouse and rat OX40 ECD, respectively. Engagement of OX40 on activated CD4+ T cells by OX40 ligand on activated dendritic cells promotes T cell survival and proliferation, prolongs the immune response, and enhances the number of cells making the transition from effector to memory T cells. OX40 signal transduction includes binding TNF receptor-associated factors (TRAFs), and activating NF kappa B and PI3 kinase to enhance expression of cytokines, antiapoptotic Bcl-2 family members, survivin and the chemokine receptor CXCR5. CXCR5 promotes T cell migration to germinal centers to deliver B cell help. Studies using knockout or transgenic mice, and agonistic or blocking antibodies, show that OX40/OX40L interaction is critical for establishing or reactivating memory T cells and breaking immune tolerance. Blockade of OX40 engagement is efficacious in animal models of allergic airway inflammation, graft-versus-host disease and autoimmune disease.

Typical data

pg/ml	O.D.		Average	Corrected
0.00	0.0321	0.0339	0.0330
46.88	0.1994	0.1959	0.1977	0.1647
93.75	0.3374	0.3551	0.3463	0.3133
187.50	0.6134	0.6371	0.6253	0.5923
375.00	1.1660	1.1100	1.1380	1.1050
750.00	2.1060	1.9760	2.0410	2.0080
1500.00	3.5420	3.4640	3.5030	3.4700
3000.00	4.4496	4.6015	4.5256	4.4926

Product Features

Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
Development protocols are provided to guide further assay optimization
Assay can be customized to your specific needs
Economical alternative to complete kits

Kit Content

Capture Antibody
Detection Antibody
Recombinant Standard
Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

ARExSet^®Ancillary Reagent Kit (5 plates): containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and assay buffer.

96 well microplates: AREX Biosciences, Catalog # DSEP01.

Plate Sealers: AREX Biosciences, Catalog # DSSF01.

Coating Buffer: 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2-7.4, 0.2 μm filtered . AREX Biosciences, Catalog # DSCB01.

Blocking Buffer: AREX Biosciences, Catalog # DSBB01.

Wash Buffer: 0.05% Tween 20 in PBS, pH 7.2-7.4. AREX Biosciences, Catalog # DSWB01.

Assay Buffer: 0.5%BSA, 0.05%Tween20, PBS Solution.AREX Biosciences, Catalog # DSAB01

Substrate Solution: Tetramethylbenzidine. AREX Biosciences, Catalog # DSTS01.

Stop Solution: 0.5 mol/ml H₂SO₄. AREX Biosciences, Catalog # DSSS01.

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