- Allow product to equilibrate to room temperature (25°C) prior to use.
- Add stop solution to plate well. Equal volumes of TMB microwell substrate and stop solution should be used. We recommend using 100 µL of TMB substrate and 100 µL of Stop Solution for TMB Substrates per well. The stopped TMB reaction product is stable for 1 hour.
- Read absorbance at 450 nm within 60 minutes. For best results, sample absorbance should be monitored and stopped before values exceed 2.0 OD units.
Mosialou I, Shikhel S, Liu JM, Maurizi A, Luo N, He Z, Huang Y, Zong H, Friedman RA, Barasch J, Lanzano P, Deng L, Leibel RL, Rubin M, Nicholas T, Chung W, Zeltser LM, Williams KW, Pessin JE, Kousteni S. MC4R-dependent suppression of appetite by bone-derived lipocalin 2. Nature. 2017. 543(7645): 385-390. Abstract.
"The reaction was terminated by addition of TMB stop solution (Immunochemistry Technologies) and absorbance was read at 450nm in a Fluostar Omega (BMG Labtech) microplate reader."
Boraschi-Diaz I, Tauer JT, El Rifai O, Guillemette D, Lefebvre G, Rauch F, Ferron M, Komarova SV. Metabolic phenotype in the mouse model of osteogenesis imperfecta. J. Endocrinol. 2017 Sep;234(3):279-289. doi: 10.1530/JOE-17-0335. Epub 2017 Jul 17. Abstract.
" – ... 2010b). Briefly, Antibody Coating Buffer (CB1), ELISA Wash Buffer (WB1), 120 General Blocker Buffer (BB1), General Assay Diluent (AD1) and Stop Solution for TMB 121 (STOP1) were all obtained from ImmunoChemistry Technologies. 1-Step™ Ultra TMB ELISA 122 ... "