Anti-UBB monoclonal antibody, clone 4D23

Reduced levels of cellular ubiquitin (Ub) pools due to disruption of the polyubiquitin gene Ubb lead to dysregulation of neural stem cell (NSC) differentiation and impaired neuronal maturation in cells isolated from Ubb(-/-) mouse embryonic brains. However, it is currently unknown whether Ub is required for the specific stage of neuronal development or whether it plays a pleiotropic role throughout the process. To answer this question, we aimed to downregulate Ubb expression temporally during neuronal development, which could not be achieved in Ubb(-/-) cells. Therefore, we exploited lentivirus-mediated knockdown (KD) of Ubb at different stages of neuronal development, and investigated their phenotypes. Here, we report the outcome of Ubb KD on two independent culture days in vitro (DIV): DIV1 and DIV7. We observed that NSCs did not differentiate properly via Ubb KD on DIV1, but the maturation of already differentiated neurons was intact via Ubb KD on DIV7. Intriguingly, Ubb KD activated Notch signaling when it had been suppressed, but exerted no effect when it had already been activated. Therefore, our study suggests that Ub plays a pivotal role in NSC differentiation to suppress Notch signaling, but not in the subsequent maturation stages of neurons that had already been differentiated.

We present a mobile laser lithography station for 3D structuring by microscopic two-photon polymerization. For structuring the Coherent Vitara UBB titanium: sapphire femtosecond laser is used, which has a power output of 500mW and generates pulses with a central wavelength of 810nm. The laser pulses have a tunable bandwidth from 50nm to 250nm. The pulses are temporally compressed using chirped mirrors to a minimum duration of less than 15fs at the sample. The laser power reaching the sample can be motionless controlled by a combination of a liquid crystal retarder and a polarizer within milliseconds. The sample is placed onto a microscope stage which has a movement range of 300 mu m in the X, Y and Z direction with an accuracy of 2nm. Sample imaging is possible with a microscope camera simultaneous to the structuring. The pulses are focused by a 40X microscope objective (1.3NA) onto the sample. To operate the lithography station, we developed a LabVIEW-based software which controls sample position, laser power and objective height and as well as the microscope camera. Furthermore, CAD data can be read and converted into sample position data. By combining all these components, a fully automatic structuring of a sample with sub-micrometer precision is possible.