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NF-L Recombinant Antibody [PS02-10] - Mouse IgG1 (Chimeric)

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Catalog# HA601448

NF-L Recombinant Antibody [PS02-10] - Mouse IgG1 (Chimeric)

Application

  • IHC-Fr

  • IHC-P

  • WB

Reactivity

  • Human

  • Mouse

  • Rat

Overview

Product Name

NF-L Recombinant Antibody [PS02-10] - Mouse IgG1 (Chimeric)

Antibody Type

Recombinant Chimeric Antibody

Immunogen

Recombinant protein.

Species Reactivity

Human, Mouse, Rat

Validated Applications

IHC-Fr, IHC-P, WB

Molecular Weight

Predicted band size: 62 kDa

Positive Control

Human cerebellum tissue, mouse cerebellum tissue, rat cerebellum tissue, Mouse brain tissue lysate, Rat brain tissue lysate.

Conjugation

unconjugated

Clone Number

PS02-10

Product Features

Form

Liquid

Concentration

1ug/ul

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Purification Method

Protein A affinity purified.

Application Dilution

  • IHC-Fr

  • 1:500

  • IHC-P

  • 1:2,000

  • WB

  • 1:5,000

Target

Function

Neurofilament light polypeptide, also known as neurofilament light chain, abbreviated to NF-L or Nfl and with the HGNC name NEFL is a member of the intermediate filament protein family. This protein family consists of over 50 human proteins divided into 5 major classes, the Class I and II keratins, Class III vimentin, GFAP, desmin and the others, the Class IV neurofilaments and the Class V nuclear lamins. There are four major neurofilament subunits, NF-L, NF-M, NF-H and α-internexin. These form heteropolymers which assemble to produce 10nm neurofilaments which are only expressed in neurons where they are major structural proteins, particularly concentrated in large projection axons. Axons are particularly sensitive to mechanical and metabolic compromise and as a result axonal degeneration is a significant problem in many neurological disorders. The detection of neurofilament subunits in CSF and blood has therefore become widely used as a biomarker of ongoing axonal compromise. The NF-L protein is encoded by the NEFL gene. Neurofilament light chain is a biomarker that can be measured with immunoassays in cerebrospinal fluid and plasma and reflects axonal damage in a wide variety of neurological disorders. It is a useful marker for disease monitoring in amyotrophic lateral sclerosis, multiple sclerosis, Alzheimer's disease, and more recently Huntington's disease. It is also promising marker for follow-up of patients with brain tumors. Higher levels of blood or CSF NF-L have been associated with increased mortality, as would be expected as release of this protein reflects ongoing axonal loss. Recent work performed as a collaboration between EnCor Biotechnology Inc. and the University of Florida showed that the NF-L antibodies employed in the most widely used NF-L assays are specific for cleaved forms of NF-L generated by proteolysis induced by cell death.

Background References

1. Gong L et al. Neurofilament Light Chain (NF-L) Stimulates Lipid Peroxidation to Neuronal Membrane through Microglia-Derived Ferritin Heavy Chain (FTH) Secretion. Oxid Med Cell Longev. 2022 Mar

2. Heiskanen M et al. Plasma Neurofilament Light Chain (NF-L) Is a Prognostic Biomarker for Cortical Damage Evolution but Not for Cognitive Impairment or Epileptogenesis Following Experimental TBI. Int J Mol Sci. 2022 Dec

Subcellular Location

Cell projection, axon, Cytoplasm, cytoskeleton.

UNIPROT

SwissProt: P07196 Human

SwissProt: P08551 Mouse

SwissProt: P19527 Rat

Synonyms

68 kDa neurofilament protein antibody

68kDa Neurofilament antibody

68kDa neurofilament protein antibody

CMT1F antibody

CMT2E antibody

FLJ53642 antibody

Light molecular weight neurofilament protein antibody

NEFL antibody

Neurofilament light antibody

Neurofilament light polypeptide 68kDa antibody

Expand

Images

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: cerebellum<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: cerebellum

    Sample: Frozen section

    Antibody concentration: 1/500

    Antigen retrieval: Not required

  • Application: IHC-Fr<br /><br />Species: Rat<br /><br />Site: cerebellum<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Rat

    Site: cerebellum

    Sample: Frozen section

    Antibody concentration: 1/500

    Antigen retrieval: Not required

  • Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Mouse anti-NF-L antibody (<a href="/products/HA601448" style="font-weight: bold;text-decoration: underline;">HA601448</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601448" style="font-weight: bold;text-decoration: underline;">HA601448</a>) at 1/2,000 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.

    Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Mouse anti-NF-L antibody (HA601448) at 1/2,000 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 (HA601448) at 1/2,000 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.

  • Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Mouse anti-NF-L antibody (<a href="/products/HA601448" style="font-weight: bold;text-decoration: underline;">HA601448</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601448" style="font-weight: bold;text-decoration: underline;">HA601448</a>) at 1/2,000 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.

    Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Mouse anti-NF-L antibody (HA601448) at 1/2,000 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 (HA601448) at 1/2,000 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.

  • Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Mouse anti-NF-L antibody (<a href="/products/HA601448" style="font-weight: bold;text-decoration: underline;">HA601448</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601448" style="font-weight: bold;text-decoration: underline;">HA601448</a>) at 1/2,000 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.

    Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Mouse anti-NF-L antibody (HA601448) at 1/2,000 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 (HA601448) at 1/2,000 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of NF-L on different lysates with Mouse anti-NF-L antibody (<a href="/products/HA601448" style="font-weight: bold;text-decoration: underline;">HA601448</a>) at 1/5,000 dilution.<br /><br />Lane 1: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 2: Mouse lung tissue lysate (negative) (20 µg/Lane)<br />Lane 3: Rat brain tissue lysate (20 µg/Lane)<br />Lane 4: Rat lung tissue lysate (negative) (20 µg/Lane)<br /><br />Predicted band size: 62 kDa<br />Observed band size: 62 kDa<br /><br />Exposure time: 2 minutes; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA601448" style="font-weight: bold;text-decoration: underline;">HA601448</a>) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of NF-L on different lysates with Mouse anti-NF-L antibody (HA601448) at 1/5,000 dilution.

    Lane 1: Mouse brain tissue lysate (20 µg/Lane)
    Lane 2: Mouse lung tissue lysate (negative) (20 µg/Lane)
    Lane 3: Rat brain tissue lysate (20 µg/Lane)
    Lane 4: Rat lung tissue lysate (negative) (20 µg/Lane)

    Predicted band size: 62 kDa
    Observed band size: 62 kDa

    Exposure time: 2 minutes; ECL: K1801;

    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 (HA601448) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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