Human/Mouse/Rat NF-L Antibody

Catalog #: MAB2216 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB2216
MAB2216-SP

Detection of Mouse and Rat NF‑L by Western Blot.

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Human/Mouse/Rat NF-L Antibody Summary

Species Reactivity

Human, Mouse, Rat

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

Monoclonal Mouse IgG_2B Clone # 268014

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human NF-L
Met1-Asp338
Accession # P07196

Formulation

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

Mouse brain (hypothalamus) and rat brain (hippocampus)

Immunohistochemistry

3-25 µg/mL

See below

Immunocytochemistry

8 µg/mL

Immersionfixed U251 human myeloma cell line (positive) and K562 human chronic myelogenous leukemia cell line (negative)

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Mouse and Rat NF‑L by Western Blot. 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.

Immunohistochemistry

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NF‑L in Human Spinal Cord. 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.

Immunohistochemistry

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NF‑L in Human Brain. 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.

Immunocytochemistry

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NF‑L in U251 human myeloma cell line (positive) and K562 human chronic myelogenous leukemia cell line (negative). 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Background: NF-L

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.

Long Name

Neurofilament Protein, Light Chain

Entrez Gene IDs

4747 (Human)

Alternate Names

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