Biolegend, 835613, Alexa Fluor® 488 anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated Antibody
25microg
Catalog Number: 835613
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Category
| Verified Reactivity | Human, Mouse, Rat |
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| Antibody Type | Monoclonal |
| Host Species | Mouse |
| Formulation | Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide. |
| Preparation | The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions. |
| Concentration | 0.5 mg/ml |
| Storage & Handling | The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze. |
| Application | IHC-P - Quality tested SB - Reported in the literature, not verified in house |
| Recommended Usage | Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 5.0 - 10 µg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application. * Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm. Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.View full statement regarding label licenses |
| Excitation Laser | Blue Laser (488 nm) |
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| Application Notes |
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| Additional reported applications (for the relevant formats of this clone): include ELISA Capture1-3, immunohistochemical staining on frozen tissue sections, immunofluorescence staining, and spatial biology (IBEX)6,7. Clone SMI 35 reacts with highly phosphorylated neurofilaments, as well as with low degrees of phosphorylation. It primarily reacts with neurofilament H and with neurofilament M to a lesser extent. Notes: On two dimensional gels, this antibody detects a band extending from the phosphorylated neurofilament position at 200 kD (pI 5.1) toward the non-phosphorylated position at 170 kD (pI 6.2). |
| Additional Product Notes | Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH). |
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| Application References(PubMed link indicates BioLegend citation) | Petzold A. 2013. J. Neuroimmunol. 262:(1-10). (ELISA) Lu CH, et al. 2012. PLoS One. 7:e40998. (ELISA) Steinacker P, et al. 2011. PLoS One. 8:e23600. (ELISA) Poltorak M, et al. 1993. J. Neurosci. 13:2217. (IHC-F) Petzold A, et al. 2011. Brain. 134:464. (WB) PubMed Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed |
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| RRID | AB_2750327 (BioLegend Cat. No. 835613) AB_2750328 (BioLegend Cat. No. 835614) |
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| Structure | The medium or middle NF (NF-M) runs at about 145-160 kD, and the heavy or highest NF (NF-H) runs at 200-220 kD. |
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| Distribution | Tissue Distribution: CNS, peripheral nerves and glandular cells of the prostate Cellular Distribution: Cytoskeleton, nucleus, cytosol, and mitochondrion |
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| Function | Neurofilaments are the major components of the neuronal cytoskeleton. They provide axonal support and regulate axon diameter. |
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| Interaction | Cell bodies and dendrites are generally unstained. Other cells and tissues are unreactive except for peripheral axons. |
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| Cell Type | Mature Neurons |
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| Biology Area | Cell Biology, Neuroscience, Neuroscience Cell Markers |
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| Molecular Family | Intermediate Filaments, Phospho-Proteins |
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| Antigen References | Petzold A. 2012. Mult Scler Int. 2012:217802. PubMed Gresle MM, et al. 2011. Mult Scler Int. 2011: 315406. PubMed |
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| Gene ID | 4744 |
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| UniProt | View information about Neurofilament HM NF-H NF-M Phospho on UniProt.org |
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| Clone | SMI 35 |
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| Regulatory Status | RUO |
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| Other Names | Neurofilament heavy polypeptide, NF-H, 200 kD neurofilament protein, neurofilament triplet H protein |
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| Isotype | Mouse IgG1, κ |
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