Cerebrospinal uid levels of p-tau (Ser 181) and p-tau (Thr 205) were determined using a Singulex Erenna immunoassay platform (USA) (Hastings et al. 2017 ). HT7 was adopted as capture antibody. AT270 and AT8 which separately recognized p-tau (Ser 181) and p-tau (Thr 205) served as detection antibodies. In the process, the measurement was performed in the presence of HaltR Protease and Phosphatase Inhibitor Cocktail (Thermo Fisher). Brie y, in 96-well plate, each well contained 10 mg capture beads, 50 μl Singulex assay buffer and 50 μl cerebrospinal uids (or standard peptide), which was diluted in PBS-T (0.2% Tween 20 in PBS). The plate was incubated at 25℃ for 4 hours, and was washed using a Biotek 405 TM TS microplate washer. Then, 20 μl AT270 or AT8 antibody (50 ng/ml) was added into the plate, and was incubated and shaken overnight at 25℃. The cerebrospinal uid specimen was developed according to the instruction, and the result was analyzed using Sgx link software (Singulex Erenna).
Haltr protease and phosphatase inhibitor cocktail
Manufactured by Thermo Fisher Scientific
HaltR Protease and Phosphatase Inhibitor Cocktail is a premixed solution designed to inhibit proteases and phosphatases in biological samples. It is intended for use in research applications to preserve the native state of proteins and prevent degradation during sample preparation and analysis.
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Lab products found in correlation
2 protocols using haltr protease and phosphatase inhibitor cocktail
1
Quantification of Tau Phosphorylation Biomarkers
2
Quantification of Tau Phosphorylation Biomarkers
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Cerebrospinal uid levels of p-tau (Ser 181) and p-tau (Thr 205) were determined using a Singulex Erenna immunoassay platform (USA) (Hastings et al. 2017 ). HT7 was adopted as capture antibody. AT270 and AT8 which separately recognized p-tau (Ser 181) and p-tau (Thr 205) served as detection antibodies. In the process, the measurement was performed in the presence of HaltR Protease and Phosphatase Inhibitor Cocktail (Thermo Fisher). Brie y, in 96-well plate, each well contained 10 mg capture beads, 50 μl Singulex assay buffer and 50 μl cerebrospinal uids (or standard peptide), which was diluted in PBS-T (0.2% Tween 20 in PBS). The plate was incubated at 25℃ for 4 hours, and was washed using a Biotek 405 TM TS microplate washer. Then, 20 μl AT270 or AT8 antibody (50 ng/ml) was added into the plate, and was incubated and shaken overnight at 25℃. The cerebrospinal uid specimen was developed according to the instruction, and the result was analyzed using Sgx link software (Singulex Erenna).
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