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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| B, H, M, Po, R | WB, ICC, IHC | M | Culture Supernatant | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB1615 |
| Replaces | 04-1112 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | Anti-Neurofilament 70 kDa Antibody, clone DA2 |
| Background Information | Neurofilaments are a type of intermediate filament that serve as major elements of the cytoskeleton supporting the axon cytoplasm. They are the most abundant fibrillar components of the axon, being on average 3-10 times more frequent than axonal microtubules. Neurofilaments (10nm in dia.) are built from three intertwined protofibrils which are themselves composed of two tetrameric protofilament complexs of monomeric proteins. The neurofilament triplet proteins (68/70, 160, and 200 kDa) occur in both the central and peripheral nervous system and are usually neuron specific. The 68/70 kDa NF-L protein can self-assemble into a filamentous structure, however the 160 kDa NF-M and 200 kDa NF-H proteins require the presence of the 68/70 kDa NF-L protein to co-assemble. Neuromas, ganglioneuromas, gangliogliomas, ganglioneuroblastomas and neuroblastomas stain positively for neurofilaments. Although typically restricted to neurons, neurofilaments have been detected in paragangliomas and adrenal and extra-adrenal pheochromocytomas. Carcinoids, neuroendocrine carcinomas of the skin, and oat cell carcinomas of the lung also express neurofilaments. For more neurofilament information see Nervous System Cell Type Specific Marker chart online under the CHEMICON Technical Support section. |
| Product Information | |
|---|---|
| Format | Culture Supernatant |
| Presentation | Liquid. Contains no preservative. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This Anti-Neurofilament 70 kDa Antibody, clone DA2 is validated for use in WB, IC, IH for the detection of Neurofilament 70 kDa. |
| Key Applications |
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| Application Notes | Immunoblot and immunohistochemistry (frozen sections, little reactivity on formalin-fixed paraffin embedded material). Optimal working dilutions must be determined by end user. IMMUNOHISTOCHEMISTRY PROTOCOL FOR MAB1615 This antibody has been used successfully on 30 mm, free floating, 4% paraformaldehyde fixed rat brain tissue. All steps are performed under constant agitation. Suggested protocol follows. 1) 3 x 10 minute washes in TBS (with or without 0.25% Triton). 2) Incubate for 30 minutes in TBS with 3% serum (same as host from secondary antibody). 3) Incubate primary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody) (with or without 0.25% Triton) for 2 hours at room temperature followed by 16 hours at 4°C. 4) 3 x 10 minute washes in TBS. 5) Incubate with secondary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody). 6) 3 x 10 minute washes in TBS. 7) ABC Elite (1:200 Vector Labs) in TBS. 8) 2 x 10 minute washes in TBS. 9) 1 x 10 minute wash in phosphate buffer (no saline). 10) DAB reaction with 0.06% NiCl added for intensification. 11) 2 x 10 minute washes in PBS. 12) 1 x 10 minute wash in phosphate buffer (no saline). |
| Biological Information | |
|---|---|
| Immunogen | Enzymatically dephosphorylated pig neurofilaments. |
| Clone | DA2 |
| Host | Mouse |
| Specificity | Recognizes the low molecular weight (68-70 kDa) subunit of the neurofilament triplet (NF-L), the epitope is phosphate independent. Reacts with human and higher vertebrates. |
| Isotype | IgG1 |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P07196 # Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. SIZE: 543 amino acids; 61517 Da DOMAIN: SwissProt: P07196 The extra mass and high charge density that distinguish the neurofilament proteins from all other intermediate filament proteins are due to the tailpiece extensions. This region may form a charged scaffolding structure suitable for interaction with other neuronal components or ions. PTM: O-glycosylated (By similarity). DISEASE: SwissProt: P07196 # Defects in NEFL are the cause of Charcot-Marie-Tooth disease type 1F (CMT1F) [MIM:607734]. CMT1F is a form of Charcot- Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT1 group are characterized by severely reduced nerve conduction velocities (less than 38 m/sec), segmental demyelination and remyelination with onion bulb formations on nerve biopsy, slowly progressive distal muscle atrophy and weakness, absent deep tendon reflexes, and hollow feet. CMT1F is characterized by onset in infancy or childhood (range 1 to 13 years). & Defects in NEFL are the cause of Charcot-Marie-Tooth disease type 2E (CMT2E) [MIM:607684]. CMT2E is an autosomal dominant form of Charcot-Marie-Tooth disease type 2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy. SIMILARITY: Belongs to the intermediate filament family. MISCELLANEOUS: NF-L is the most abundant of the three neurofilament proteins and, as the other nonepithelial intermediate filament proteins, it can form homopolymeric 10-nm filaments. |
| Product Usage Statements | |
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| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain at -20°C in undiluted aliquots for up to 12 months. Avoid repeated freeze-thaw cycles. |
| Packaging Information | |
|---|---|
| Material Size | 300 µL |