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Tissue/ cell preparation: human umbilical vein endothelial cells (HUVECs).
Our Abpromise guarantee covers the use of ab24577 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 5 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 72 kDa).|
|ELISA||Use a concentration of 1 - 10 µg/ml.|
|IHC-Fr||1/1000. See Abreview.|
|Flow Cyt||1/1000. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
ab24577 at 1/1000 dilution staining mouse brain tissue sections by Immunohistochemistry (Frozen sections). Mice were processed by transcardial perfusion first with saline, then 4% PF. After overnight incubation in 4% PF, brains were transfered to sucrose. Upon saturation, brains were frozen, sectioned with a cryostat, and then the sections were immediately mounted on slides. The tissue was incubated with ab24577 for 2 hours and then an Alexa Fluor ® 594 goat anti-mouse IgG was used as the secondary (red). DAPI staining is shown in blue. Images were taken with a confocal microscope in comparable cortex regions of the lesion or contralateral side in the same section. The lesion image shown is from this model of ischemia-hypoxia, with 1 hour of recovery time after injury, when endothelial cell activation is quite robust.