Neurofilament Antibodies-Check Them Out

Widely Used and Frequently Published
Our Neurofilament Antibodies are well characterized and research ready. Here is a recent publication using one of our Neurofilament Heavy Antibodies. Jennifer M. Hahn, Kelly A. Combs, Christopher M. Lloyd, Kevin L. McFarland, Steven T. Boyce, and Dorothy M. Supp. Identification of Merkel cells associated with neurons in engineered skin substitutes after grafting to full thickness wounds. PLoS One. 2019; 14(3): e0213325. Published online 2019 Mar 5. doi: 10.1371/journal.pone.0213325.

Figures: Merkel cells in grafted ESS are associated with neurons expression neurofilament heavy (NF-M) by eight weeks after grafting. Immunochemistry with antibodies against NF-H (red) and KRT20 (green) was used to localize neurons and Merkel cells, respectively, in ESS after grafting to mice. Nuclei were counterstained with DAPI (blue; B, D, F, H). Shown are cross sections of ESS at 4 weeks (A-B), 8 weeks (C-D), 12 weeks (E-F), and 14 weeks (G-H) after grafting; each row contains images of the same section. Scale bars in A is same for all images. White arrows indicate examples of NF-H-positive nerves associated with or in proximity to Merkel cells; yellow arrows indicate NF-H-positive nerves not associated with Merkel cells.

Remember. All our antibodies come with a money back guarantee. Your satisfaction is joh #1 for us. 

Neuronal Markers

New Pub References 3 Markers
We are recognized for our large catalog of neuronal markers. Our strength, in this area, includes markers designed for pain researchers.

They are widely used and frequently published. This new publication references use of our Guinea Pig Substance P, Guinea Pig PGP9.5 and Chicken NF200 of NF-Heavy. andla, Jagadeesha, Lomada, Santosh Kumara, Jianninga; Kuner, Rohinia, Bali, Kiran Kumar. miR-34c-5p functions as pronociceptive microRNA in cancer pain by targeting Cav2.3 containing calcium channels. Pain: September 2017 - Volume 158 - Issue 9 - p 1765–1779 doi: 10.1097/j.pain.0000000000000971.

Neuromics' PGP9.5 Staining of  Mouse DRGs.

.Neuromics SP and NF200 Staining of Mouse DRGSs

Immuno-fluorescence in 3-D

Potent Markers!

We are seeing more 3-D Cell Based Assays being used for Neuroscience Research. It is important researchers have the Neuron-Astroglial-Progenitor Markers required for staining in 3-D. This helps researchers determine the cell types present in their assays.

I have posted here published results. I would like to share the latest: Yiting Liu, Katherine S. Given, Danielle E. Harlow, Adeline M. Matschulat, Wendy B. Macklin, Jeffrey L. Bennett and Gregory P. Owens. Myelin-specific multiple sclerosis antibodies cause complement-dependent oligodendrocyte loss and demyelination. Acta Neuropathologica Communications Neuroscience of Disease 20175:25
DOI: 10.1186/s40478-017-0428-6

3D movie reconstructed by super-resolution structured illumination microscopy (SIM) imaging of live organotypic mouse cerebellar slices stained with MS#30 (red), then fixed and stained for MAG (blue) and NF-H (purple). MS#30 reactivity was on oligodendrocyte processes, including those contacting adjacent axons, and on myelinated MAG+ axons, outside of MAG layer. Scale bar: 2 μm. (MPG 90340 kb).

We will continue to post cool applications for our antibodies here.

Neurite Outgrowth Assays

Cells, Media and Markers

I have previously posted use of our Neurons in Live Content Assays for the study of Neurite Outgrowth: Neurons-Live Content Assays. These assays are critical for the study of repair and regeneration.

A recent publication featured several of our Neuron Markers: Serena Quarta, Bastian E. Baeumer, Nadja Scherbakov1, Manfred Andratsch, Stefan Rose-John, Georg Dechant3, Christine E. Bandtlow, and Michaela Kress: Peripheral Nerve Regeneration and NGF-Dependent Neurite Outgrowth of Adult Sensory Neurons Converge on STAT3 Phosphorylation Downstream of Neuropoietic Cytokine Receptor gp130. The Journal of Neuroscience, 24 September 2014, 34(39): 13222-13233; doi: 10.1523/JNEUROSCI.1209-13.2014.
Live labeling of neuron cultures: After 20 or 48 h, neurons were live-labeled with α-gp130 antibody diluted in cold TNB medium for 30 min on ice. After washing, neurons were incubated with the secondary antibody diluted in cold TNB medium for 30 min and washed with PBS. Cells were fixed either with 4% PFA for 20 min at room temperature (RT) or with methanol at −20°C for 2 min. After permeabilization with 0.01% TX-100 (Pierce) unspecific binding was blocked for 30 min with 10% normal goat serum (Sigma-Aldrich) in PBS. Cells were incubated with the first antibody for 1 h, washed three times for 10 min with PBS and incubated with the appropriate secondary antibody for 30 min, counterstained with 4′, 6-diamidino-2-phenylindole (1:10,000; Sigma-Aldrich) and embedded in Mowiol (Calbiochem). As primary antibodies, α-gp130 (1:50; Neuromics), α-β-III-tubulin clone TuJ-1 (1:1000; R&D Systems), and α-neurofilament-H (α-NF-H; 1:200; Neuromics) were used. Secondary antibodies used were α-goat AlexaFluor 594 (1:1000; Invitrogen), chicken α-mouse AlexaFluor 594 or donkey α-mouse AlexaFluor 488 (1:1000; Invitrogen), and goat α-chicken AlexaFluor 568 (1:10,000; Invitrogen) for fluorescence microcopy.

Images: Reduced density of TuJ-1+ nerve endings in the epidermis in SNS-gp130−/− mice after lesion. A, Representative cross sections of hindpaw glabrous skin of naive and 12 dpl gp130fl/fl and SNS-gp130−/− mice stained with the pan neuronal marker TuJ-1. The dotted line indicates the border between dermis and epidermis. Scale bar, 40 μm. B, Quantification of the total number of TuJ-1+ fibers (NE) per 1000 μm2 of epidermal area shows a significant decrease in density in SNS-gp130−/− mice after lesion compared with control animals (*p < 0.05; n = 4 for each group). Data are presented as mean ± SEM and analyzed by Mann–Whitney U test. C, 3D reconstruction of the deeper layer of the dermis shows fewer nerve bundles in SNS-gp130−/− dermis compared with controls. D, No NF-H+ proprioceptive fibers were detectable in the epidermis of gp130fl/fl animals at 12 dpl. Scale bar, 40 μm.

If you want to learn more about our Neuron-Glial-Astrocyte based assay solutions do not hesitate to contact me (612-801-1007) or pshuster@neuromics.com. Pete Shuster, Owner and CEO, Neuromics.

Arthritic Pain, Nerve Sprouting and Neuroma Formation

Our friends, Drs Mantyh and Jimenez Andrade, have published important findings on the potential root cuases of age related Arthritic Pain: Juan M Jimenez-Andrade and Patrick W Mantyh. Sensory and sympathetic nerve fibers undergo sprouting and neuroma formation in the painful arthritic joint of geriatric mice. Note: our Chicken Polyclonal NF200 Antibody was used to label primary afferent sensory nerve fibers.

Overview: Introduction: Although the prevalence of arthritis dramatically increases with age, the great majority of preclinical studies concerning the mechanisms that drive arthritic joint pain have been performed in young animals. One mechanism hypothesized to contribute to arthritic pain is ectopic nerve sprouting; however, neuroplasticity is generally thought to be greater in young versus old nerves. Here we explore whether sensory and sympathetic nerve fibers can undergo a significant ectopic nerve remodeling in the painful arthritic knee joint of geriatric mice.

In other words, there is a clear root reason that the pain arthritis sufferers experience increases overtime. Here's some related data. Notice the increase in labeled nerves:

Images. Sensory nerve fiber sprouting and formation of neuroma-like structures in the painful geriatric arthritic knee joint. Schematic of a frontal view of a cross-sectioned mouse knee joint (A). The red square illustrates the synovial region from which the confocal images were obtained. Representative confocal images of calcitonin gene-related peptide (CGRP+), neurofilament 200 kDa (NF200+) sensory nerve fibers (yellow/orange) and growth associated protein (GAP43; marker of fibers undergo regeneration, yellow/orange) and DAPI labeled nuclei (blue) in knee joint sections (20 μm thick) of vehicle-injected (B,D,F) and CFA-injected (C,E,G ) mice. In vehicle-injected mice, a low level, regular pattern of innervation by CGRP+ and NF200+ fibers is observed in the synovial space of the knee joint. Twenty-eight days following the initial CFA injection, a significant number of CGRP+ and NF200+ nerve fibers have sprouted and have a disorganized appearance as compared to vehicle-injected mice. Note that, CGRP+ and NF200+ sprouted nerve fibers are localized in the synovium and are not observed in the meniscus of the joint. Furthermore, there was formation of neuroma-like structures in the synovium of the geriatric mice injected with CFA (G).

Related Reagents: Neuronal-Glial Markers-Astrocytes, Glia, Microglia, Olidogodendrocytes, Progenitors and Schwann Cell Markers.

ENS Development Markers

This is an excellent study on the developing Enteric Nervous Stem (ENS). This system is often referred to as the "second brain".

Despite this, neurogenesis has been much less studied in the ENS than in the brain. Understanding how neurons are formed in the gut is the foundation for finding cures for ENS disorders. The key finding here is that the ability of 5-HT4 receptors to unmask a regulation of enteric neurogenesis in adult animals suggests that the mature ENS is capable of an unexpected degree of plasticity (potentially good new for discovering therapies for ENS related disorders).

Min-Tsai Liu, Yung-Hui Kuan, Jingwen Wang, René Hen, and Michael D. Gershon. 5-HT4 Receptor-Mediated Neuroprotection and Neurogenesis in the Enteric Nervous System of Adult Mice. The Journal of Neuroscience, August 5, 2009, 29(31):9683-9699; doi:10.1523/JNEUROSCI.1145-09.2009.

More details @ http://www.anxietyinsights.info/serotonin_the_gut_and_neurogenesis.htm

On a side note, this publication is rich with excellent ICC and Western Blot images of a variety of Neurogenesis Markers. This include referencing of 4 of our markers:

GFAP-CH22102-ICC Dilution 1:2000
Musashi-1-RA14128-ICC Dilution 1:100 WB Dilution 1:1000
Neurofilament NF-H-CH22104-ICC Dilution 1:2000
S100B-RA25022-ICC Dilution 1;1000