Aptagen, LLC
VR11 (ID# 9090)
DNA
TNFα
Protein
7 nM (reported value)
A 96-well ELISA microtiter plate (BD Falcon) was coated overnight at 4 °C with 100 μL of human TNFα (10 μg/mL) prepared in coating buffer (0.2 M carbonate/bicarbonate, pH 9.4). The plate was then washed 3 times with PBS-T (0.1 M phosphate, 0.15 M sodium chloride, pH 7.0 containing 0.05% Tween-20) and blocked overnight at 4 °C in 150 μL of blocking solution (PBS-T + 1% (w/v) BSA). The plate was washed 3 times with PBS-T and incubated overnight at 4 °C with 100 μL (10 μg/mL) of 5′ biotinylated aptamers dissolved in PBS, 0.005% (v/v) Tween-20. The plates were washed and incubated for 1 h at 4 °C with 100 μL of streptavidin-HRP (1:2000). Plates were then read on a plate reader at 450 nm following washing and dispensing of 100 μL of TMB substrate (1 TMB tablet (Sigma-Aldrich Inc.) dissolved in 1 mL of DMSO in 10 mL of 0.05 M phosphate citrate, pH 5.0 and 2 μL sodium peroxide) per well. Color development was stopped by adding 50 μL of 1 M sulphuric acid per well.
37 °C°C
NA If the oligo is a known aptamer sequence: For binding studies, perform a refolding protocol to ensure proper function (i.e. binding to antigen or target). Refer to the aptamer reference source for the appropriate refolding parameters and binding conditions. Note: it is unknown whether aptamer functions properly without refolding.
dTpdGpdGpdTpdGpdGpdApdTpdGpdGpdCpdGpdCpdApdGpdTpdCpdGpdGpdCpdGpdApdCpdApdAp
25
7788.1 g/mole
244400 L/(mole·cm)
Note: Information on this aptamer oligo was obtained from the literature and hasn't been validated by Aptagen.
Orava EW, Jarvik N, Shek YL, Sidhu SS, Gariépy J. A short DNA aptamer that recognizes TNFα and blocks its activity in vitro. ACS Chem Biol. 2013 Jan 18;8(1):170-8. doi: 10.1021/cb3003557. Epub 2012 Oct 24. PMID: 23046187.
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