Ultra tmb elisa reagent

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Ultra TMB-ELISA reagent is a highly sensitive substrate for the detection and quantification of horseradish peroxidase (HRP) in enzyme-linked immunosorbent assay (ELISA) applications. The reagent provides a colorimetric signal that can be measured spectrophotometrically.

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Lab products found in correlation

Nunc immuno 96 microwell solid plates, by Thermo Fisher Scientific (1 mentions) Multiskan microplate photometer, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

ELISAs (35 citations) Ultra-TMB (22 citations) Ultra TMB-ELISA (11 citations) TMB reagent (10 citations) Ultra-TMB reagent (5 citations) 1-Step Ultra TMB-ELISA reagent (2 citations)

2 protocols using ultra tmb elisa reagent

Bacterial OMV-Based ELAA for GN6 Aptamer

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For GN6 ELAA using bacterial OMVs, Nunc-Immuno 96 MicroWell solid plates (Thermo Scientific, USA) were used to immobilize bacterial OMVs in Tris buffer. After incubating OMVs at various concentrations in the plate overnight at 4 °C, the plate was washed twice and blocked using 2% BSA-Tris buffer for 2 h. After blocking, 20 pmol of GN6 aptamer and N40 control were separately added and incubated for 1 h. After washing 4 times, streptavidin-Poly HRP conjugate (Pierce, USA) was added and incubated for 30 min. After thoroughly washing 5 times in Tris buffer with 0.05% Tween-20, Ultra TMB-ELISA reagent (Thermo Scientific, USA) was added. After 15 min, 1 M sulfuric acid as stop solution was added. Absorbance at 450 nm was measured using Multiskan microplate photometer (Thermo Scientific, USA). The measured values were analyzed using non-linear regression fit model of SigmaPlot 12.0.

Shin H.S., Gedi V., Kim J.K, & Lee D.K. (2019). Detection of Gram-negative bacterial outer membrane vesicles using DNA aptamers. Scientific Reports, 9, 13167.

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Quantitative Aptamer Binding Assay

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Selected and control phNA aptamer sequences were synthesized from a FITC-labelled DNA primer (or 2’O-methyl primer) and a DNA template and PAGE-purified as described previously. For binding assays the DNA portion of the phNA aptamer was hybridized to its complementary sequence at a 1:1 ratio. All sequences were hybridized and assayed in phosphate-buffered saline (PBS) and 0.1% Tween. Aptamer clones were bound to Streptavidin wells (or Neutravidin, or Goat IgG controls) for 1 hour, then unbound aptamers were washed three times with buffer, and an anti-FITC Fab-HRP (Roche) was bound at 0.15 units mL^-1, and unbound Fab was removed. After washing the wells three times with wash buffer, the assay was developed using Ultra-TMB ELISA reagent (Thermo). Peroxidase reactions were stopped with 1M H₂SO₄ before absorbance measurements (450 nm).

Arangundy-Franklin S., Taylor A.I., Porebski B.T., Genna V., Peak-Chew S., Vaisman A., Woodgate R., Orozco M, & Holliger P. (2019). Encoded synthesis and evolution of alkyl-phosphonate nucleic acids: a synthetic genetic polymer with an uncharged backbone chemistry. Nature chemistry, 11(6), 533-542.

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