Immunohistochemistry (IHC)
(Figure 2. IHC analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).NEFL/NF-L was detected in paraffin-embedded section of mouse spinal cord tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NEFL/NF-L Antibody (MBS1753527) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Rabbit NEFL/NF-L Polyclonal Antibody | anti-NEFL antibody
Anti-NEFL/NF-L Antibody
IHC-P: 2-5ug/ml|Mouse, Rat|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).NEFL/NF-L was detected in paraffin-embedded section of mouse spinal cord tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NEFL/NF-L Antibody (MBS1753527) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).NEFL/NF-L was detected in paraffin-embedded section of mouse spinal cord tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NEFL/NF-L Antibody (MBS1753527) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of 293T cells using anti-NEFL/NF-L antibody (MBS1753527).Overlay histogram showing 293T cells stained with MBS1753527 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NEFL/NF-L Antibody (MBS1753527, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of 293T cells using anti-NEFL/NF-L antibody (MBS1753527).Overlay histogram showing 293T cells stained with MBS1753527 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NEFL/NF-L Antibody (MBS1753527, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Western Blot (WB)
(Figure 1. Western blot analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human SH-SY5Y whole cell lysatesLane 2: rat brain tissue lysatesLane 3: mouse brain tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-NEFL/NF-L antigen affinity purified polyclonal antibody (Catalog # MBS1753527) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for NEFL/NF-L at approximately 72KD. The expected band size for NEFL/NF-L is at 72KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human SH-SY5Y whole cell lysatesLane 2: rat brain tissue lysatesLane 3: mouse brain tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-NEFL/NF-L antigen affinity purified polyclonal antibody (Catalog # MBS1753527) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for NEFL/NF-L at approximately 72KD. The expected band size for NEFL/NF-L is at 72KD. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).NEFL/NF-L was detected in paraffin-embedded section of rat spinal cord tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NEFL/NF-L Antibody (MBS1753527) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).NEFL/NF-L was detected in paraffin-embedded section of rat spinal cord tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NEFL/NF-L Antibody (MBS1753527) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).NEFL/NF-L was detected in paraffin-embedded section of mouse spinal cord tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NEFL/NF-L Antibody (MBS1753527) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of NEFL/NF-L using anti-NEFL/NF-L antibody (MBS1753527).NEFL/NF-L was detected in paraffin-embedded section of mouse spinal cord tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NEFL/NF-L Antibody (MBS1753527) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
2. Hurst, J., Flavell, D., Julien, J. -P., Meijer, D., Mushynski, W., Grosveld, F. The human neurofilament gene (NEFL) is located on the short arm of chromosome 8. Cytogenet. Cell Genet. 45: 30-32, 1987.
3. Leung, C. L., Nagan, N., Graham, T. H., Liem, R. K. H. A novel duplication/insertion mutation of NEFL in a patient with Charcot-Marie-Tooth disease. Am. J. Med. Genet. 140A: 1021-1025, 2006.
NCBI and Uniprot Product Information
NCBI Description
Neurofilaments are type IV intermediate filament heteropolymers composed of light, medium, and heavy chains. Neurofilaments comprise the axoskeleton and they functionally maintain the neuronal caliber. They may also play a role in intracellular transport to axons and dendrites. This gene encodes the light chain neurofilament protein. Mutations in this gene cause Charcot-Marie-Tooth disease types 1F (CMT1F) and 2E (CMT2E), disorders of the peripheral nervous system that are characterized by distinct neuropathies. A pseudogene has been identified on chromosome Y. [provided by RefSeq, Oct 2008]
Uniprot Description
NFL: one of the three (L, M, and H) intermediate filament proteins that form neurofilaments. Neurofilaments are involved in the maintenance of neuronal caliber. NF-L is the most abundant of the three neurofilament proteins. Defects cause Charcot-Marie-Tooth disease type 1F (CMT1F).
Protein type: Cytoskeletal
Chromosomal Location of Human Ortholog: 8p21
Cellular Component: TSC1-TSC2 complex; growth cone; axon; cytoplasm; neurofilament; cytosol
Molecular Function: protein C-terminus binding; protein binding, bridging; identical protein binding; protein domain specific binding; protein binding; phospholipase binding; structural constituent of cytoskeleton
Biological Process: protein polymerization; response to peptide hormone stimulus; neurofilament bundle assembly; response to toxin; intermediate filament organization; microtubule cytoskeleton organization and biogenesis; retrograde axon cargo transport; axon regeneration in the peripheral nervous system; synaptic transmission; axon transport of mitochondrion; regulation of axon diameter; positive regulation of axonogenesis; negative regulation of neuron apoptosis; anterograde axon cargo transport; neuromuscular process controlling balance; locomotion; neurite morphogenesis; response to corticosterone stimulus
Disease: Charcot-marie-tooth Disease, Axonal, Type 2e; Charcot-marie-tooth Disease, Demyelinating, Type 1f
Research Articles on NEFL
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Product Notes
The NEFL nefl (Catalog #AAA1753527) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-NEFL/NF-L Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's NEFL/NF-L can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human, Mouse, Rat| IHC-P: 2-5ug/ml|Mouse, Rat| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the NEFL nefl for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "NEFL/NF-L, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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