Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH13-10] to Human tPA antibody (Capture) (HA725115) and Recombinant Human tPA protein (HA211026) as the standard. The reference range value is 140-100,000 pg/mL.
ELISA
Use at an assay dependent concentration.
Target
Function
Tissue-type plasminogen activator, short name tPA, is a protein that facilitates the breakdown of blood clots. It acts as an enzyme to convert plasminogen into its active form plasmin, the major enzyme responsible for clot breakdown. It is a serine protease (EC 3.4.21.68) found on endothelial cells lining the blood vessels. Human tPA is encoded by the PLAT gene, and has a molecular weight of ~70 kDa in the single-chain form. tPA can be manufactured using recombinant biotechnology techniques, producing types of recombinant tissue plasminogen activator (rtPA) such as alteplase, reteplase, and tenecteplase. These drugs are used in clinical medicine to treat embolic or thrombotic stroke, but they are contraindicated and dangerous in cases of hemorrhagic stroke and head trauma. The antidote for tPA in case of toxicity is aminocaproic acid.
Background References
1. Wang R et al. Neutrophil extracellular traps promote tPA-induced brain hemorrhage via cGAS in mice with stroke. Blood. 2021 Jul
2. Cai Y et al. FUNDC1-dependent mitophagy induced by tPA protects neurons against cerebral ischemia-reperfusion injury. Redox Biol. 2021 Jan
Sandwich ELISA analysis of Human tPA matched pair antibodies
Capture: HA725115, Human tPA Rabbit mAb [PSH13-10] Detector: HA725116, Human tPA Rabbit mAb [PSH13-11]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725115) diluted in carbonate/bicarbonate buffer, at a concentration of 0.5 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human tPA protein (HA211026) starting from 100,000 pg/ml to 0 pg/ml and detect antibody (HA725116, HRP, 0.5 µg/ml) for 1 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
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