Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
![Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)](https://www.abcam.com/ps/products/7/ab7794/Images/ab7794-412722-anti-160-kd-neurofilament-medium-antibody-nf-09-neuronal-marker-western-blot-human-knockout-lysate.jpg "Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)")
Key features and details
- Mouse monoclonal [NF-09] to 160 kD Neurofilament Medium - Neuronal Marker
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Cow, Human, Pig
- Isotype: IgG2a
Overview
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Product name
Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker
See all 160 kD Neurofilament Medium primary antibodies -
Description
Mouse monoclonal [NF-09] to 160 kD Neurofilament Medium - Neuronal Marker -
Host species
Mouse -
Specificity
This antibody reacts with both phosphorylated and non-phosphorylated forms of medium neurofilament protein (160 kDa) of various species. -
Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human, Pig
Predicted to work with: a wide range of other species, Mammals
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Immunogen
Pellet of pig brain cold stable proteins after depolymerization of microtubules.
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Positive control
- WB: HEK-293 and A549 whole cell lysates ICC/IF: Dental pulp stem, Neuro2A, PC12 and human nerve cells
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.097% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
NF-09 -
Isotype
IgG2a -
Light chain type
unknown -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
Applications
Our Abpromise guarantee covers the use of ab7794 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/1000. | |
| ICC/IF | Use at an assay dependent concentration. |
Target
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Relevance
Neurofilaments are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called neurofilament light (NF-L), neurofilament medium (NF-M) and neurofilament heavy (NF-H). Neurofilament medium runs on SDS-PAGE gels in the range 145-170 kDa, with some variation in different species. Antibodies to this protein are useful to identify neurons and their processes in tissue sections and in tissue culture. Neurofilament medium can also be useful in studies of neurofilament accumulations seen in many neurological diseases, such as Lou Gehrig's disease or Alzheimer's disease. -
Cellular localization
Cytoplasm -
Database links
- Entrez Gene: 281347 Cow
- Entrez Gene: 4741 Human
- Entrez Gene: 18040 Mouse
- Entrez Gene: 24588 Rat
- Omim: 162250 Human
- SwissProt: O77788 Cow
- SwissProt: P07197 Human
- SwissProt: P08553 Mouse
see all -
Alternative names
- 150kDa medium antibody
- NEF3 antibody
- NEFM antibody
see all
Images
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Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)All lanes : Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : NEFM knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Observed band size: 150 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab7794 observed at 150 kDa. Red - loading control ab181602 observed at 36 kDa.
ab7794 Anti-160 kD Neurofilament Medium antibody [NF-09] was shown to specifically react with 160 kD Neurofilament Medium in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266741 (knockout cell lysate ab257103) was used. Wild-type and 160 kD Neurofilament Medium knockout samples were subjected to SDS-PAGE. ab7794 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)](https://www.abcam.com/ps/products/7/ab7794/Images/ab7794-13254-anti-160-kd-neurofilament-medium-antibody-nf-09-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)Image from Ferro F et al., PLoS One. 2012;7(7):e41774. Epub 2012 Jul 23. Fig 3.; doi:10.1371/journal.pone.0041774; July 23, 2012, PLoS ONE 7(7): e41774.Immunofluorescence analysis of 1 month neuronal-differentiated dental pulp stem cells, staining 160 kD Neurofilament Medium with ab7794.
Cells were fixed with paraformaldehyde and incubated with primary antibody (1/600). A FITC-conjugated anti-mouse IgG (1/375) was used as the secondary antibody. -
![Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)](https://www.abcam.com/ps/products/7/ab7794/Images/ab7794-345715-Anti-160-kD-Neurofilament-Medium-antibody-NF09-western-blot-human.jpg)
Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)All lanes : Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : NEFM knockout HEK-293 whole cell lysate
Lane 3 : A549 whole cell lysate
Lysates/proteins at 20 µg per lane.
Observed band size: 150 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab7794 observed at 150 kDa. Red - loading control, ab52866, observed at 50 kDa.
ab7794 was shown to specifically react with NEFM (Neurofilament) in wild-type HEK-293 cells as signal was lost in NEFM knockout cells. Wild-type and NEFM knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab7794 and ab52866 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)](https://www.abcam.com/ps/products/7/ab7794/Images/ab7794-279998-anti-160-kd-neurofilament-medium-antibody-nf-09-western-blot.jpg)
Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)Western blotting analysis of neurofilament medium protein in porcine brain lysate (reducing conditions) by mouse monoclonal NF-09.
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![Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)](https://www.abcam.com/ps/products/7/ab7794/Images/ab7794-13255-anti-160-kd-neurofilament-medium-antibody-nf-09-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)ab7794 staining Neurofilament medium protein in mouse Neuro2A cells by ICC/IF. -
![Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)](https://www.abcam.com/ps/products/7/ab7794/Images/ab7794-13256-anti-160-kd-neurofilament-medium-antibody-nf-09-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)ICC/IF image of ab7794 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7794, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor© 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43æM.
References (33)
ab7794 has been referenced in 33 publications.
- Ma L et al. Sennoside A restores colonic barrier function through protecting colon enterocytes from ROS-induced mitochondrial damage in diet-induced obese mice. Biochem Biophys Res Commun 526:519-524 (2020). PubMed: 32245617
- Fan YN et al. Characterization of Group I Metabotropic Glutamate Receptors in Rat and Human Adrenal Glands. Front Physiol 11:401 (2020). PubMed: 32536873
- Jung W et al. Wire Electrodes Embedded in Artificial Conduit for Long-term Monitoring of the Peripheral Nerve Signal. Micromachines (Basel) 10:N/A (2019). PubMed: 30871203
- Huang CW et al. Multipotent vascular stem cells contribute to neurovascular regeneration of peripheral nerve. Stem Cell Res Ther 10:234 (2019). PubMed: 31376835
- Jin J et al. Igf1R/InsR function is required for axon extension and corpus callosum formation. PLoS One 14:e0219362 (2019). PubMed: 31318893
