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, ELISA, ICC/IF, IHC-Fr, IHC-P, IHC (PFA fixed), ICC
- Knockout validated
- Reacts with: Mouse, Rat, Cow, Cat, 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, ELISA, ICC/IF, IHC-Fr, IHC-P, IHC (PFA fixed), ICCmore details -
Species reactivity
Reacts with: Mouse, Rat, Cow, Cat, 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: Dental pulp stem, Neuro2A, PC12 and human nerve cells
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, 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
The Abpromise guarantee
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 |
Use a concentration of 1 - 2 µg/ml.
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| ELISA |
Use at an assay dependent concentration.
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| ICC/IF |
Use at an assay dependent concentration.
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| IHC-Fr | (1) |
Use at an assay dependent concentration.
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| IHC-P | (1) |
Use at an assay dependent concentration.
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| IHC (PFA fixed) |
Use at an assay dependent concentration.
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| ICC |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
Use a concentration of 1 - 2 µg/ml. |
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ELISA
Use at an assay dependent concentration. |
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ICC/IF
Use at an assay dependent concentration. |
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IHC-Fr
Use at an assay dependent concentration. |
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IHC-P
Use at an assay dependent concentration. |
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IHC (PFA fixed)
Use at an assay dependent concentration. |
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ICC
Use at an assay dependent concentration. |
Target
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Function
Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. -
Sequence similarities
Belongs to the intermediate filament family. -
Post-translational
modificationsThere are a number of repeats of the tripeptide K-S-P, NFM is phosphorylated on a number of the serines in this motif. It is thought that phosphorylation of NFM results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber.
Phosphorylation seems to play a major role in the functioning of the larger neurofilament polypeptides (NF-M and NF-H), the levels of phosphorylation being altered developmentally and coincidentally with a change in the neurofilament function.
Phosphorylated in the head and rod regions by the PKC kinase PKN1, leading to the inhibition of polymerization. - Information by UniProt
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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
- 160 kDa neurofilament protein antibody
- NEF3 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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![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)
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 - 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 - 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.
Datasheets and documents
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SDS download
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Datasheet download
References (38)
ab7794 has been referenced in 38 publications.
- Choi SW et al. Mammalian Cochlear Hair Cell Imaging Using Optical Coherence Tomography (OCT): A Preliminary Study. J Int Adv Otol 17:46-51 (2021). PubMed: 33605221
- Jia J et al. Transcriptional factor FoxM1-activated microRNA-335-3p maintains the self-renewal of neural stem cells by inhibiting p53 signaling pathway via Fmr1. Stem Cell Res Ther 12:169 (2021). PubMed: 33691791
- 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
- Wang D et al. Valproic acid-labeled chitosan nanoparticles promote recovery of neuronal injury after spinal cord injury. Aging (Albany NY) 12:8953-8967 (2020). PubMed: 32463791