NF-L Polyclonal antibody

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  WB analysis of mouse brain tissue using 12998-1-AP

  WB result of 12998-1-AP.

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  WB analysis using 12998-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 12998-1-AP (NF-L antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse brain using 12998-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 12998-1-AP (NF-L antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

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  WB analysis of Neuro-2a using 12998-1-AP

  Neuro-2a cells were subjected to SDS PAGE followed by western blot with 12998-1-AP (NF-L antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  IP experiment of SH-SY5Y using 12998-1-AP

  IP result of anti-NF-L (IP:12998-1-AP, 4ug; Detection:12998-1-AP 1:40000) with SH-SY5Y cells lysate 880 ug.

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  IHC staining of mouse brain using 12998-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 12998-1-AP (NF-L antibody) at dilution of 1:2000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse brain using 12998-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 12998-1-AP (NF-L antibody) at dilution of 1:2000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse brain using 12998-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 12998-1-AP (NF-L antibody) at dilution of 1:2000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse brain using 12998-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using NF-L antibody (12998-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  IF Staining of SH-SY5Y using 12998-1-AP

  Immunofluorescent analysis of (4% PFA) fixed SH-SY5Y cells using NF-L antibody (12998-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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  IF Staining of SH-SY5Y using 12998-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed SH-SY5Y cells using NF-L antibody (12998-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  FC experiment of SH-SY5Y using 12998-1-AP

  1x10^6 SH-SY5Y cells were intracellularly stained with 0.4 ug Anti-Human NF-L (12998-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

NF-L Polyclonal Antibody for WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ELISA

Cat No. 12998-1-AP

Publications(26)

Reviews (1)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ELISA, IHC-IF

Synonyms

NEFL, NFL, NF68, neurofilament

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Tested Applications

• Positive WB detected in: SH-SY5Y cells, Neuro-2a cells, mouse brain tissue, C6 cells
• Positive IP detected in: SH-SY5Y cells
• Positive IHC detected in: 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
• Positive IF/ICC detected in: SH-SY5Y cells
• Positive FC (Intra) detected in: SH-SY5Y cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:16000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• 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 13 publications below
• IHC: See 2 publications below
• IF: See 10 publications below
• IP: See 1 publications below
• ELISA: See 1 publications below

Product Information

12998-1-AP targets NF-L in WB, IHC, IF/ICC, IF-P, FC (Intra), IP, ELISA, IHC-IF applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: NF-L fusion protein Ag3667
• Full Name: neurofilament, light polypeptide
• Calculated Molecular Weight: 543 aa, 62 kDa
• Observed Molecular Weight: 65-68 kDa, 140-160 kDa
• GenBank Accession Number: BC039237
• Gene Symbol: NF-L
• Gene ID (NCBI): 4747
• RRID: AB_10597388
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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. It has a very weak reaction to NEFM.

Publications

Species	Application	Title
mouse	WB	Mol Cell; Mixed Lineage Kinase Domain-like Protein MLKL Breaks Down Myelin following Nerve Injury.; Authors - Zhengxin Ying
mouse	WB	Mater Today Bio; Injectable hyaluronic acid hydrogel loaded with BMSC and NGF for traumatic brain injury treatment.; Authors - Luyu Wang
mouse	WB	Mater Today Bio; In situ forming and biocompatible hyaluronic acid hydrogel with reactive oxygen species-scavenging activity to improve traumatic brain injury repair by suppressing oxidative stress and neuroinflammation.; Authors - Dan Zhang
rat	WB,IF	Adv Sci (Weinh); An Important Function of Petrosiol E in Inducing the Differentiation of Neuronal Progenitors and in Protecting Them against Oxidative Stress.; Authors - Jing Liu
human	ELISA	Anal Chem; Electrochemical Biosensor for Point-of-Care Testing of Low-Abundance Biomarkers of Neurological Diseases; Authors - Ziyue Huang
human	IHC	Acta Neuropathol Commun; Multiparametric grading of glaucoma severity by histopathology can enable post-mortem substratification of disease state; Authors - Chuanxi Xiang

Reviews

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

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FH

Max (Verified Customer) (02-24-2022)

strong and specific labeling of NfL in hamster brain lysate on western blot

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: Hamster brain lysate