NF-L Monoclonal antibody

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  WB analysis using 60189-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60189-1-Ig (NF-L antibody) at dilution of 1:100000 incubated at room temperature for 1.5 hours.

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  IHC staining of rat brain using 60189-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 60189-1-Ig (NF-L antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of rat brain using 60189-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 60189-1-Ig (NF-L antibody) at dilution of 1:10000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse brain using 60189-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 60189-1-Ig (NF-L antibody) at dilution of 1:200 (under 0x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human brain using 60189-1-Ig

  Immunohistochemical analysis of paraffin-embedded human brain tissue slide using 60189-1-Ig (NF-L antibody) at dilution of 1:400 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse brain using 60189-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 60189-1-Ig (NF-L antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  IF Staining of rat brain using 60189-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed rat brain tissue using NF-L antibody (60189-1-Ig, Clone: 5C12G4 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  FC experiment of PC-12 using 60189-1-Ig

  1X10^6 PC-12 cells were intracellularly stained with 0.4 ug Anti-Human NF-L (60189-1-Ig, Clone:5C12G4) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

NF-L Monoclonal Antibody for WB, IHC, IF-P, FC (Intra), ELISA

Cat No. 60189-1-Ig

Clone No.5C12G4

Publications(4)

Reviews (1)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, pig

Applications

WB, IHC, IF-P, FC (Intra), ELISA

Synonyms

NEFL, NFL, NF68, neurofilament, 5C12G4

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Tested Applications

• Positive WB detected in: PC-12 cells, human brain tissue, pig brain tissue, rat brain tissue, mouse brain tissue
• Positive IHC detected in: rat brain tissue, human brain tissue, mouse brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: mouse brain tissue, rat brain tissue
• Positive FC (Intra) detected in: PC-12 cells

Recommended dilution

• Western Blot (WB): WB : 1:20000-1:100000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 2 publications below
• IF: See 2 publications below

Product Information

60189-1-Ig targets NF-L in WB, IHC, IF-P, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat, pig samples.

• Tested Reactivity: human, mouse, rat, pig
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: NF-L fusion protein Ag15178
• Full Name: neurofilament, light polypeptide
• Calculated Molecular Weight: 543 aa, 62 kDa
• Observed Molecular Weight: 65 kDa
• GenBank Accession Number: BC039237
• Gene Symbol: NF-L
• Gene ID (NCBI): 4747
• RRID: AB_10860271
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P07196
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

NEFL, also named as NF68 and NF-L, belongs to the intermediate filament family. Neurofilaments are the 10 nm intermediate filaments found specifically in neurons. They are a major component of the cell's cytoskeleton, and provide support for normal axonal radial growth. Neurofilaments usually contain three intermediate filament proteins: L, M, and H, which are involved in the maintenance of neuronal caliber. The names given to the three major neurofilament subunits are based upon the apparent molecular weight of the mammalian subunits on SDS-PAGE: NF-L, 65-68 kDa; NF-M,145-160 kDa and NF-H, 200-220 kDa.This antibody is specific to NEFL.

Publications

Species	Application	Title
mouse	IF	Cell Rep Med; Cholinergic signaling via muscarinic M1 receptor confers resistance to docetaxel in prostate cancer; Authors - Jing Wang
human,mouse	WB,IF	ACS Nano; Microarray Chip-Based High-Throughput Screening of Neurofilament Light Chain Self-Assembling Peptide for Noninvasive Monitoring of Alzheimer's Disease; Authors - Ying Zhou
rat	WB	J Stroke Cerebrovasc Dis; Edaravone dexborneol regulates γ-aminobutyric acid transaminase in rats with acute intracerebral hemorrhage; Authors - Rui Yang
human	FC (Intra)	J Neuroinflammation; CX3CR1+/UCHL1+ microglial extracellular vesicles in blood: a potential biomarker for multiple sclerosis; Authors - Jing Duan

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Reyes (Verified Customer) (10-18-2024)

NF-L worked perfectly on my FFPE human brain sections.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:100
• Cell Tissue Type: Human Brain Cortex

NF-L Antibody Immunofluorescence validation (1:100 dilution) in Human Brain Cortex (Cat no:60189-1-Ig)