B6 mice were injected intravenously with 250 μg of Alexa Fluor 488–conjugated human fibrinogen (ThermoFisher) ~30 min before mTBI. As shown in Supplementary Movie 2 , fibrinogen leaked from injured vessels immediately following mTBI. Fluorescent fibrinogen that leaked into the extravascular spaces would remain there unless cleared or degraded. Remaining extravascular fibrinogen at 7 d after injury was quantified by collection of a 100-μm z-stack through the thinned skull bone by two-photon microscopy. This scan was performed to visualize the un-cleared extravascular fibrinogen deposits. Fibrinogen fluorescence was quantified using Imaris as described as described below (‘Two photon image analysis’).
Alexa fluor 488 conjugated human fibrinogen
Manufactured by Thermo Fisher Scientific
Alexa Fluor 488–conjugated human fibrinogen is a fluorescently labeled protein used for research purposes. It is produced by conjugating the Alexa Fluor 488 dye to human fibrinogen, a blood plasma glycoprotein involved in blood clotting. This product can be utilized in various biochemical and cell-based applications that require the detection or visualization of fibrinogen.
Automatically generated - may contain errors
2 protocols using alexa fluor 488 conjugated human fibrinogen
1
Visualizing Extravascular Fibrinogen Post-mTBI
2
Hypochlorous Acid and Antioxidant Interactions
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In this study, we used NaOCl solution as a “hypochlorous acid” (pKa 7.45). This refers to ~50% ionized mixture of HOCl and ClO−, existing at physiological pH (as “HOCl” throughout the text). Before each experiment, the exact concentration of HOCl was assessed by spectrophotometric measurement using a 292 nm wavelength and a molar extinction coefficient of 350 M−1cm−1. Quercetin, resveratrol, epigallocatechin gallate, and sodium hypochlorite solution were from Sigma-Aldrich (Merck). Collagen (type I) was from Chrono-Log Corp. Alexa Fluor 488-conjugated human fibrinogen was from Thermo Fisher Scientific. Alteplase (recombinant human tissue plasminogen activator) was from Boehringer Ingelheim.
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