|Immunopanning: Protocols and Antibodies
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Welcome to the latest edition of Biosensis e-antibody solutions. Many researchers are purifiying and isolating specific subpopulations of cells (stem cells, neurons) from tissue or blood extracts for analysis or culture. While Fluorescence Activated Cell Sorting (FACS) is commonly used, Immunopanning has a number of advantages, particularly in providing a very gentle approach to neuronal cell purification.
Two detailed protocols (and one update) from 3 leading research laboratories are provided to help researchers with this important technique.
The first (from Germany and Australia) is from Nature Protocols and gives details of how to isolate Motor Neurons from embryonic spinal cord: Wiese S. et al, (2010) Isolation and enrichment of embryonic mouse motoneurons from the lumbar spinal cord of individual mouse embryos.
(You can see an update of this protocol here:
Klausmeyer et al, (2015) Isolation and Culture of Spinal Cord Motor Neurons.
The second is from a leading Stanford researcher in Cold Spring Harbor Protocols.
Ben Barres. (2014) Designing and Troubleshooting Immunopanning Protocols for Purifying Neural Cells. http://www.ncbi.nlm.nih.gov/pubmed/?term=25447277
This publication also provides references to several other well-established protocols for a variety of neuronal and glial subpopulations. For example, here is a useful Protocol for immunopanning sub-populations of neurons from the rat Dorsal Root Ganglion:
Zuchero JB, (2014). http://www.ncbi.nlm.nih.gov/pubmed/?term=25086011
Above figures are images of stem cell derived neurons that have stained positive for p75 Nerve Growth Factor (CD271) taken directly from the Moxi Flow. The Moxi Flow outputs p75NGFR negative versus p75NGFR+ (12.2%) with the counts displayed in the black text boxes. The left image shows a scatter plot of fluorescence versus cell volume (diameter), and the right image displays the diameter and fluorescence only histograms.
Moxi Flow is an instrument owned and distributed by Orflo Technologies. Images are courtesy of Orflo Technologies.
Biosensis monoclonal antibodies ideal for immunopanning
Biosensis provides numerous monoclonal antibodies that are ideal for immunopanning.
p75NTR Monoclonals for mouse, rat and human neurons and stem cells.
Clone MLR2 for mouse neurons and stem cells. (Catalog# M-009-100), (Catalog# M-018-100-FT)
Clone MC192 for rat neurons and stem cells. (Catalog# M-006-100), (Catalog# M-016-100-FT)
Clone ME20.4 for human neurons and stem cells. (Catalog# M-011-100), (Catalog# M-017-100-FT)
A selection of other monoclonal antibodies suitable for immunopanning primary sensory neurons, as well as various central neuronal subpopulations are also available. Here is a selection:
Capsaicin Receptor monoclonals and Trk Receptor antibodies
Mouse monoclonal antibody to rat capsaicin receptor. (Catalog# M-1714-100)
Mouse monoclonal antibody to Tyrosine Kinase Receptor A (TrkA). (Catalog# M-1719-100)
Mouse monoclonal antibody to Tyrosine Kinase Receptor B (TrkB). New, please enquire.
Mouse monoclonal antibody to Tyrosine Kinase Receptor C (TrkC). New, please enquire.
Mouse monoclonal antibody to CD11b (clone OX42). (Catalog# M-1325-100).
Mouse monoclonal antibody to Orexin Receptor type 2 (Clone 1E3). (Catalog# M-840-100).
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