Anti-160 kD Neurofilament Medium antibody [EP2460] - Neuronal Marker KO Tested (ab92539)

Knockout Tested Rabbit Recombinant Monoclonal NFM antibody. Suitable for WB, IHC-P, IHC-Fr, ICC/IF and reacts with Human samples. Cited in 4 publications.

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Images

Western blot - Anti-160 kD Neurofilament Medium antibody [EP2460] - Neuronal Marker (AB92539), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	IHC-Fr	ICC/IF
Human	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

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Target data

See full target information NEFM

Function

Neurofilaments usually contain three intermediate filament proteins: NEFL, NEFM, and NEFH which are involved in the maintenance of neuronal caliber. May additionally cooperate with the neuronal intermediate filament proteins PRPH and INA to form neuronal filamentous networks (By similarity).

Alternative names

NEF3, NFM, NEFM, Neurofilament medium polypeptide, NF-M, 160 kDa neurofilament protein, Neurofilament 3, Neurofilament triplet M protein

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal NFM antibody. Suitable for WB, IHC-P, IHC-Fr, ICC/IF and reacts with Human samples. Cited in 4 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EP2460
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The 160 kD Neurofilament Medium also known as NEFM NF-M or NEF-M belongs to the family of neurofilament proteins. This protein is an important structural component of neurons particularly in the axonal cytoskeleton. Neurofilament Medium has a molecular mass of around 160 kilodaltons (kD). It shows high levels of expression in neuronal tissues contributing to the maintenance of axonal caliber. Commonly research recognizes it alongside other neurofilament proteins emphasizing its role in providing structural integrity to neurons.

Biological function summary

Neurofilament Medium participates in the formation of a stable network of neurofilaments within neurons. Acting as a part of the intermediate filament protein family it forms a complex with other neurofilaments such as NF-L and NF-H. This complex supports neuron structure and plays an important role in axonal transport. The heteropolymeric nature of neurofilaments contributes to their mechanical stability facilitating essential neuronal functions by aligning neurofilaments along the axon.

Pathways

Neurofilament Medium integrates into the cytoskeletal arrangement pathways that govern axonal transport and stability. It plays a significant role in the neuronal transport pathway associating with other proteins such as dynein and kinesin responsible for the motor functions along axons. Also it relates to the MAP kinase pathway where phosphorylation events modulate its assembly and disassembly dynamics in response to cellular signals.

Associated diseases and disorders

Neurofilament Medium closely relates to neurodegenerative conditions such as amyotrophic lateral sclerosis (ALS) and Alzheimer's disease. Aberrant phosphorylation or aggregation of NEFM can disrupt neuronal function leading to pathology. In ALS the abnormal accumulation of neurofilaments correlates with motor neuron degeneration. In Alzheimer's its interaction with tau protein poses significant interest as altered states of either could exacerbate neurofibrillary tangles worsening cognitive decline.

Product promise

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12 product images

Western blot - Anti-160 kD Neurofilament Medium antibody [EP2460] - Neuronal Marker (ab92539)
160 kD Neurofilament Medium Western blot staining using rabbit Anti-160 kD Neurofilament Medium antibody
Anti-NEFM antibody [EP2460] (ab92539) staining at 1/100000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab92539 was shown to bind specifically to NEFM. A band was observed at 140-160 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in NEFM knockout cell line. To generate this image, wild-type and NEFM knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween$^®$ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-160 kD Neurofilament Medium antibody [EP2460] - Neuronal Marker (ab92539) at 1/100000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: NEFM knockout HEK-293T cell lysate at 20 µg
Lane 3: SK-N-BE cell lysate at 20 µg
Lane 4: A549 cell lysate at 20 µg
Lane 5: SH-SY5Y cell lysate at 20 µg
Secondary
Lanes 1 - 5: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 5: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 102 kDa
Observed band size: 140-160 kDa
Western blot - Anti-160 kD Neurofilament Medium antibody [EP2460] - Neuronal Marker (ab92539)
160 kD Neurofilament Medium Western blot staining using rabbit Anti-160 kD Neurofilament Medium antibody
All lanes: Western blot - Anti-160 kD Neurofilament Medium antibody [EP2460] - Neuronal Marker (ab92539) at 1/100000 dilution
Lane 1: 293T cell lysate at 10 µg
Lane 2: SH-SY5Y cell lysate at 10 µg
Secondary
All lanes: HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution
Predicted band size: 102 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
Immunohistochemical analysis of paraffin-embedded Human liver sections labelling 160 kD Neurofilament Medium with ab92539 at 1/5000 dilution, followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) system. Counterstained with Hematoxylin.
Negative control: no staining on human liver. The section was incubated with ab92539 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
Immunohistochemical analysis of paraffin-embedded Human astrocytoma sections labelling 160 kD Neurofilament Medium with ab92539 at 1/5000 dilution, followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) system. Counterstained with Hematoxylin.
Positive staining on human astrocytoma. The section was incubated with ab92539 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
ab92539 showing negative staining in Normal breast tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
ab92539 showing negative staining in Normal human liver tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
ab92539 showing negative staining in Normal human colon tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
ab92539 showing negative staining in Normal human tonsil tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
ab92539, at 1/250 dilution, staining 160 kD Neurofilament Medium in formalin-fixed, paraffin-embedded Human glioma tissue by immunohistochemistry.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
ab92539, at 1/250 dilution, staining 160 kD Neurofilament Medium in formalin-fixed, paraffin-embedded Human normal brain tissue by immunohistochemistry.
Immunohistochemistry (Frozen sections) - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
Immunohistochemistry image of Neurofilament 160 kDa staining in a section of frozen normal human cerebral cortex*.
The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab92539 at 1/500 dilution. The secondary antibody was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.The section was then mounted using Fluoromount^®.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539)
ab92539 stained in SHSY5Y cells. Cells were fixed with 100% methanol (5 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with the antibody ab92539 at 0.5µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed ab150087 (colored red) and Donkey Anti-Sheep IgG H&L (Alexa Fluor® 488) ab150177 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
This product gave a positive signal in 4% formaldehyde (10 min) fixed SHSY5Y cells under the same testing conditions.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).