NF-L Antibody (PS02-10)

Western Blot: NF-L Antibody (PS02-10) [NBP3-32650] - Western blot analysis of NF-L on different lysates with Rabbit anti-NF-L antibody (NBP3-32650) at 1/1,000 dilution. Lane 1: Mouse brain tissue lysate Lane 2: Rat brain tissue lysate Lane 3: Mouse hippocampus tissue lysate Lane 4: Rat hippocampus tissue lysate Lane 5: Human liver tissue lysate (negative) Lane 6: Mouse liver tissue lysate (negative) Lane 7: Rat liver tissue lysate (negative) Lysates/proteins at 20 ug/Lane. Predicted band size: 62 kDa Observed band size: 68 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32650) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: NF-L Antibody (PS02-10) [NBP3-32650] - Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-NF-L antibody (NBP3-32650) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32650) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemistry: NF-L Antibody (PS02-10) [NBP3-32650] - Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling NF-L with Rabbit anti-NF-L antibody (NBP3-32650) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (NBP3-32650, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Immunocytochemistry/ Immunofluorescence: NF-L Antibody (PS02-10) [NBP3-32650] - Immunocytochemistry analysis of SH-SY5Y cells labeling NF-L with Rabbit anti-NF-L antibody (NBP3-32650) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NF-L antibody (NBP3-32650) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF, IHC
• Clone: PS02-10
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

NF-L Antibody (PS02-10) Summary

• Description: Novus Biologicals Rabbit NF-L Antibody (PS02-10) (NBP3-32650) is a recombinant monoclonal antibody validated for use in IHC, WB and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Recombinant protein. (Uniprot: P07196)
• Localization: Cell projection, axon, Cytoplasm, cytoskeleton.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: NEFL
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:1000
  • Western Blot 1:1000
• Theoretical MW: 62 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.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.1% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for NF-L Antibody (PS02-10)

• 68 kDa neurofilament protein
• CMT2E
• NEFL
• neurofilament protein, light chain
• neurofilament subunit NF-L
• Neurofilament triplet L protein
• neurofilament, light polypeptide
• neurofilament-light
• NF68
• NF68FLJ53642
• NFL
• NF-L
• NFLlight polypeptide 68kDa

Background

Neurofilaments are 10nm intermediate filament proteins located in vertebrate neurons, which regulate axonal diameter. They are composed predominantly of the three major neurofilament proteins: NF-Light, NF-Medium and NF-Heavy. NF-L is the light or low molecular weight polypeptide, and it runs on SDS-PAGE gels at about 68kDa, with some size variation in across species.

Antibodies to NF-L may be used to identify NF-L in neurons and to study neuronal processes in tissue sections and/or culture. NF-L antibodies can also be used to study microfilament accumulations seen in many neurological diseases, such as Lou Geri's disease or Alzheimer's disease.

Limitations

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.

Bioinformatics

• Gene Symbol: NEFL