NF-L Antibody (268014) [Unconjugated]

Western blot shows lysates of mouse brain (hypothalamus) and rat brain (hippocampus). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat NF‑L Monoclonal Antibody (Catalog # MAB2216) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018) . A specific band was detected for NF‑L at approximately 74 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

NF-L was detected in immersion fixed paraffin-embedded sections of human spinal cord using 25 µg/mL Mouse Anti-Human NF-L Monoclonal Antibody (Catalog # MAB2216) overnight at 4 °C. Tissue was stained with the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counter-stained with hematoxylin (blue). Specific labeling was localized to the cytoplasm in motor neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

NF-L was detected in immersion fixed paraffin-embedded sections of human brain (substantia nigra) using Mouse Anti-Human NF-L Monoclonal Antibody (Catalog # MAB2216) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

NF‑L was detected in immersion fixed U251 human myeloma cell line (positive) and K562 human chronic myelogenous leukemia cell line (negative) using Mouse Anti-Human/Mouse/Rat NF‑L Monoclonal Antibody (Catalog # MAB2216) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: Hu, Mu, Rt
• Applications: WB, IHC, ICC/IF
• Clone: 268014
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

NF-L Antibody (268014) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human NF-L
  Met1-Asp338
  Accession # P07196
• Specificity: Detects human NF-L in direct ELISAs and human, mouse and rat NF-L in Western blots. In direct ELISAs and Western blots, no cross‑reactivity with recombinant human (rh) NF‑H or rhNF‑M is observed.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: NEFL
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8 ug/mL
  • Immunohistochemistry 3-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for NF-L Antibody (268014) [Unconjugated]

• 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

NF-L is a 68 kDa light chain cytoskeletal intermediate filament protein that is expressed in neurons. It associates with the 125 kDa NF-M and the 200 kDa NF-H to form neurofilaments.

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
• Uniprot:
  • P07196()