Antibodies, Proteins, Kits and Reagents for Life Science

294 835 ₸ Product size

100 µl 294 835 ₸

Order now and get it on Tuesday March 02, 2021

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20020] to Neurofilament heavy polypeptide
Suitable for: IHC-Fr, ICC/IF, IHC-P, WB
Reacts with: Mouse, Rat, Human

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 & Species

Application	Species
ICC/IF	Mouse
IHC-Fr	Mouse Rat
IHC-P	Mouse Rat Human
WB	Mouse Rat Human

See all applications and species data

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

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)

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