Research in the Laboratory of Developmental Neurobiology, headed by Dr. Herbert Geller, is focused on understanding how extracellular signals provide guidance cues to axons. Our primary focus is on signals derived from the extracellular matrix, especially proteoglycans, as well as responses of growing neurons to biophysical properties of their environment, such as substrate stiffness. Our overall goal is to improve recovery of function following brain or spinal cord injury. This involves use of all methods of modern biology, including light and electron microscopy, including multi-photon and superresolution microscopy, cell biology, including proteomics, constrol of gene expression using CRISPR/Cas systems, and animal models.
Western Blotting for Chondroitin Sulfate Proteoglycans
This protocol was developed by Dr. Panpan Yu, Associate Professor, Guangdong–Hongkong–Macau Institute of CNS Regeneration, Jinan University, Guangzhou, China when she was a Research Fellow in the lab. Please contact her at: firstname.lastname@example.org: for further help.
Protein samples are separated by SDS-PAGE on a 6% polyacrylamide gel under reducing conditions.
Electroblot the SDS-PAGE-separated samples onto the PVDF membrane using semi-dry or wet transfer device. (Note: Reduce the methanol concentration in transfer buffer to 10 % instead of 20%, to facilitate transfer of high molecular weight proteins such as CSPGs).
Block the membrane in blocking buffer (5% milk in PBS-Tween) at room temperature for 1 h or overnight at 4°C on the shaker.
Incubate the membrane overnight at 4°C on the shaker with primary antibody CS-56 (1:500, Sigma) diluted in 5% milk in PBS-Tween; incubate for additional 2 h the following day at room temperature.
Wash the membrane 3 times for 10 min each with PBS-Tween.
Incubate the membrane with HRP-conjugated goat anti-mouse IgM secondary antibody at room temperature for 35 min.
Wash the membrane 6 times for 5 min each with PBS-Tween.
Detect the signals using chemiluminescent methods. (Note: use high sensitive chemiluminescent reagents)
Meet the Team
Herbert Geller, Ph.D.
Herbert Geller graduated with a Ph.D.in Biomedical Engineering from Case Western Reserve University and was Postdoctoral Fellow in Physiology at the University of Rochester. He was a Professor in the Departments of Pharmacology and Neurology at Robert Wood Johnson Medical School in New Jersey before moving to NIH in 2001. Dr. Geller has authored or co-authored more than 150 papers, and he sits on the editorial boards of the International Journal of Developmental Neuroscience and the Journal of Neuroscience Methods. At NIH, he has been a mentor in the Biomedical Engineering Summer Internship Program. His former trainees have gone onto careers in Academia, Industry and Government. Dr. Geller is a member of the Society for Neuroscience and a Fellow of the American Association for the Advancement of Science. In 2018, Dr. Geller was awarded the Bernice Grafstein Award for Outstanding Accomplishments in Mentoring from the Society for Neuroscience.