NF-M Antibody

Novus Biologicals | Catalog # NB300-133

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Chicken, Equine

Cited:

Human, Mouse, Rat

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
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Product Specifications

Immunogen

Recombinant fusion protein containing the extreme C-terminal segment of rat NF-M, amino acids 549-845

Marker

Neuronal Marker

Specificity

Specifically recognizes the evolutionarily conserved extreme C-terminal region of Neurofilament Medium (~145-170kDa).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

160 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for NF-M Antibody

Knockout Validated: NF-M Antibody [NB300-133]

Western Blot: NF-M Antibody [NB300-133]

Western Blot: NF-M Antibody [NB300-133] - Analysis lysates of HeLa human cervical epithelial carcinoma parental cell line and NF-M knockout (KO) HeLa cell line. PVDF membrane was probed with 1:5,000 of Rabbit Anti-Human NF-M Polyclonal Antibody (Catalog # NB300-133) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for NF-M at approximately 155 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.
Immunocytochemistry/ Immunofluorescence: NF-M Antibody [NB300-133]

Immunocytochemistry/ Immunofluorescence: NF-M Antibody [NB300-133]

Immunocytochemistry/Immunofluorescence: NF-M Antibody [NB300-133] - Analysis of rat cerebellum section stained with rabbit pAb to NF-M, NB300-133, dilution 1:2,000 in red, and costained with mouse mAb to GAP43, dilution 1:2,000 in green. Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45uM, and free-floating sections were stained with the above antibodies. NB300-133 antibody strongly labels neuronal processes throughout the cerebellum, while the GAP43 antibody stains predominantly synaptic regios in the molecular layer.
Western Blot: NF-M Antibody [NB300-133]

Western Blot: NF-M Antibody [NB300-133]

Western Blot: NF-M Antibody [NB300-133] - Analysis of neuronal tissue lysates using rabbit pAb to NF-M, NB300-133, dilution 1:2,000 in green: [1] protein standard (red), [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord, [6] pig brain, [7] pig spinal cord. Strong bands at 145kDa correspond to rodent NF-M molecules, while the NF-M of pig and other larger mammals including humans run at about 160kDa.
Immunohistochemistry-Frozen: NF-M Antibody [NB300-133]

Immunohistochemistry-Frozen: NF-M Antibody [NB300-133]

Immunohistochemistry-Frozen: NF-M Antibody [NB300-133] - Staining of cerebral cortex section from Rat with NF-M antibody (red) at 1:5000 dilution. This immunostaining reveals the perikarya of pyramidal neurons and dendrites as well as axons surrounding the neurons. The green channel shows staining with a monoclonal antibody to beta-adrendergic receptor kinase 1.
Western Blot: NF-M Antibody [NB300-133]

Western Blot: NF-M Antibody [NB300-133]

Western Blot: NF-M Antibody [NB300-133] - Analysis of 160kDa Neurofilament Medium expression on whole rat cerebellum homogenate using NB300-133.
Immunocytochemistry/ Immunofluorescence: NF-M Antibody [NB300-133]

Immunocytochemistry/ Immunofluorescence: NF-M Antibody [NB300-133]

Immunocytochemistry/Immunofluorescence: NF-M Antibody [NB300-133] - Mixed neuron/glia cultures stained with NB300-133 (red) and NBP2-25141 (green), Novus' mouse monoclonal antibody to Fox1, an mRNA binding protein closely related to Fox3/NeuN. The RPCA-NF-M antibody stains axonal, dendritic and perikaryal profiles of neurons cleanly and specifically. Like antibody to Fox3/NeuN, the Fox1 antibody binds to the nuclei of neurons only. DNA is shown in blue with the DAPI stain.

Applications for NF-M Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:1000-1:2500

Immunohistochemistry

1:1000-1:2500

Immunohistochemistry-Frozen

1:1000-1:2500

Immunohistochemistry-Paraffin

1:1000-1:2500

Western Blot

1:1000-1:5000
Application Notes
This 160kDa Neurofilament Medium Antibody is useful for WB, ICC/IF, IHC-P, and IHC-F. In WB a band can be seen around 145kDa corresponding to rodent Neurofilament Medium. Human and bovine Neurofilament Medium run a little slower and can be seen at about 160kDa. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Supplied as serum

Preservative

0.035% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: NF-M

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 is the middle or medium neurofilament subunit and runs on SDS-PAGE gels in the range 145-170kDa, with some variation in different species. Antibodies to Neurofilament Medium are useful to identify this protein and 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.

Long Name

Neurofilament Protein, Medium Chain

Alternate Names

NEF3, NEFM, NFM

Entrez Gene IDs

4741 (Human); 18040 (Mouse); 24588 (Rat)

Gene Symbol

NEFM

UniProt

Additional NF-M Products

Product Documents for NF-M Antibody

Certificate of Analysis

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Product Specific Notices for NF-M Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

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