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Immunohistochemical staining of tissue sections was performed using the Envision System HRP mouse (Dako, Glostrup, Denmark).
The slides were then treated with hydrogen peroxide, followed by incubation with the primary and secondary antibodies, a streptavidin-biotin complex, an amplification reagent, streptavidin-peroxidase and substrate-chromogen solution using the Envision system according to the manufacturers' protocol (DAKO).
CX3CR1 expression was normalized among samples using the POLR2A expression level and was represented as fold of CX3CR1 expression relative to the THP-1 cell line, arbitrarily set at 1. Immunohistochemical staining of tonsil tissue sections was performed using the Envision System HRP and alkaline phosphatase (AP) mouse (DAKO), as reported [47].
Detection was performed using the Envision system (DakoCytomation, Glostrup, Denmark).
Immunostaining was performed using the Envision System with diaminobenzidine (Dako, Glostrup, Denmark).
The localization of the bound monoclonal antibodies was detected using the Envision System (DakoCytomation).
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CD133 detection was performed by using the EnVision system-HRP (Dako, Glostrup, Denmark) in a DakoCytomation Autostainer platform, according to the manufacturer's instructions.
For the immunohistochemical detection of ICAM-1, we used the EnVision System from Dako.
Immunohistochemistry staining was performed using the EnVision™ system (EnVision system, Dako, USA) [ 22].
Slides were the incubated with relevant secondary antibodies for 1 hour, i.e. either secondary goat-anti-mouse Alexa Fluor® 594 or goat-anti-rabbit Alexa Fluor® 594 antibodies (Invitrogen, Eugene, Oregon, USA) or using the Envision+ system (Dako, Denmark) according to the manufacturers instructions.
Antibody detection was performed using the EnVision™ system.
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