Recombinant Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Product name

Anti-Neurofilament heavy polypeptide antibody [EPR20020]
See all Neurofilament heavy polypeptide primary antibodies
Description

Rabbit monoclonal [EPR20020] to Neurofilament heavy polypeptide
Host species

Rabbit
Tested applications

Suitable for: IHC-Fr, ICC/IF, IHC-P, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Mouse and rat cerebellum and brain lysates; Neuro-2a and SH-SY5Y whole cell lysates; Human midbrain lysate. IHC-P: Human cerebrum and cerebellum tissues; Mouse cerebellum tissue; Rat cerebral cortex tissue. IHC-Fr: Mouse hippocampus tissue; Rat cerebral cortex tissue. ICC/IF: Neuro-2a cells.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20020
Isotype

IgG
Research areas
- Neuroscience
- Development

Alternative Versions
- Anti-Neurofilament heavy polypeptide antibody [EPR20020] - BSA and Azide free (ab223139)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Neurofilament heavy polypeptide protein (ab158986)
Related Products
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab207176 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
IHC-Fr		1/500.
ICC/IF		1/100.
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/1000. Detects a band of approximately 200 - 210 kDa (predicted molecular weight: 112 kDa).

Notes
IHC-Fr 1/500.
ICC/IF 1/100.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 200 - 210 kDa (predicted molecular weight: 112 kDa).

Function

Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. NF-H has an important function in mature axons that is not subserved by the two smaller NF proteins.
Involvement in disease

Defects in NEFH are a cause of susceptibility to amyotrophic lateral sclerosis (ALS) [MIM:105400]. ALS is a neurodegenerative disorder affecting upper and lower motor neurons, and resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology is likely to be multifactorial, involving both genetic and environmental factors.
Sequence similarities

Belongs to the intermediate filament family.
Post-translational
modifications

There are a number of repeats of the tripeptide K-S-P, NFH is phosphorylated on a number of the serines in this motif. It is thought that phosphorylation of NFH 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 coincident with a change in the neurofilament function.
Phosphorylated in the Head and Rod regions by the PKC kinase PKN1, leading to inhibit polymerization.
Target information above from: UniProt accession P12036 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 4744 Human
- Entrez Gene: 380684 Mouse
- Entrez Gene: 24587 Rat
- Omim: 162230 Human
- SwissProt: P12036 Human
- SwissProt: P19246 Mouse
- SwissProt: P16884 Rat
- Unigene: 198760 Human
- Unigene: 298283 Mouse
- Unigene: 108194 Rat
see all
Alternative names
- 200 kDa neurofilament protein antibody
- CMT2CC antibody
- Nefh antibody
- Neurofilament heavy polypeptide 200kDa antibody
- Neurofilament heavy polypeptide antibody
- Neurofilament triplet H protein antibody
- NF H antibody
- NF-H antibody
- NFH antibody
- NFH_HUMAN antibody
see all

Immunohistochemistry (Frozen sections) - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus tissue labeling Neurofilament heavy polypeptide with ab207176 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Positive staining on mouse hippocampus is observed.

The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Neurofilament heavy polypeptide with ab207176 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on the normal Human cerebrum tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling Neurofilament heavy polypeptide with ab207176 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on Neuro-2a cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Western blot - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Lanes 1 & 3 : Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176) at 1/1000 dilution
Lanes 2 & 4 : Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176) at 1/2000 dilution

Lane 1 : Mouse cerebellum lysate
Lane 2 : Mouse brain lysate
Lane 3 : Rat cerebellum lysate
Lane 4 : Rat brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 112 kDa
Observed band size: 200 - 210 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1/2: 3 seconds; Lane 3/4: 1 second.

The MW observed is consistent with the literature: PMID: 25374587; PMID: 144556.
Western blot - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176) at 1/1000 dilution + Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 112 kDa
Observed band size: 200 - 210 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The MW observed is consistent with the literature: PMID: 25374587; PMID: 144556.
Western blot - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Lane 1 : Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176) at 1/1000 dilution
Lane 2 : Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176) at 1/2000 dilution

Lane 1 : Human midbrain lysate
Lane 2 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lane 1 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 112 kDa
Observed band size: 200 - 210 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 1 minute; Lane 2: 30 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling Neurofilament heavy polypeptide with ab207176 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on Human cerebellum is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling Neurofilament heavy polypeptide with ab207176 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on mouse cerebellum is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling Neurofilament heavy polypeptide with ab207176 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on neurons of the rat cerebral cortex is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Neurofilament heavy polypeptide with ab207176 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative control: No staining on human kidney.

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Frozen sections) - Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebral cortex tissue labeling Neurofilament heavy polypeptide with ab207176 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Positive staining on rat cerebral cortex is observed.

The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Anti-Neurofilament heavy polypeptide antibody [EPR20020] (ab207176)

Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab207176? Please let us know so that we can cite the reference in this datasheet.

ab207176 has been referenced in 12 publications.

Luo Y et al. Tocilizumab promotes repair of spinal cord injury by facilitating the restoration of tight junctions between vascular endothelial cells. Fluids Barriers CNS 20:1 (2023). PubMed: 36624478
Li Z et al. SU16f inhibits fibrotic scar formation and facilitates axon regeneration and locomotor function recovery after spinal cord injury by blocking the PDGFRβ pathway. J Neuroinflammation 19:95 (2022). PubMed: 35429978
Xu Z et al. Positive interaction between GPER and β-alanine in the dorsal root ganglion uncovers potential mechanisms: mediating continuous neuronal sensitization and neuroinflammation responses in neuropathic pain. J Neuroinflammation 19:164 (2022). PubMed: 35729568
Yao F et al. Imatinib inhibits pericyte-fibroblast transition and inflammation and promotes axon regeneration by blocking the PDGF-BB/PDGFRβ pathway in spinal cord injury. Inflamm Regen 42:44 (2022). PubMed: 36163271
Liu R et al. In Vivo Clonal Analysis Reveals Development Heterogeneity of Oligodendrocyte Precursor Cells Derived from Distinct Germinal Zones. Adv Sci (Weinh) 8:e2102274 (2021). PubMed: 34396711

View all Publications for this product

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Post-translationalmodifications

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (12)

Customer reviews and Q&As

Post-translational
modifications