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NF-M Recombinant Rabbit Monoclonal Antibody [JM11-20] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751854

NF-M Recombinant Rabbit Monoclonal Antibody [JM11-20] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IHC-Fr

  • IF-Tissue

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Predicted reactivity

  • Cynomolgus monkey

Conjugation

  • unconjugated

Overview

Product Name

NF-M Recombinant Rabbit Monoclonal Antibody [JM11-20] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human NEFM aa 651-692 / 916.

Species Reactivity

Human, Mouse, Rat (Predicted: Cynomolgus monkey)

Predicted species support after-sales service

Validated Applications

WB, IHC-P, IHC-Fr, IF-Tissue

Target Molecular Weight

Predicted band size: 102 kDa

Positive Control

Mouse cerebellum tissue, rat cerebellum tissue, human cerebellum tissue, 293T cell lysate, Rat brain tissue lysate, Mouse brain tissue lysate.

Conjugation

unconjugated

Clone Number

JM11-20

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:2,000

  • IHC-Fr

  • 1:500

  • IF-Tissue

  • 1:500

Target

Function

Neurofilament-M (NF-M), for neurofilament medium polypeptide, a member of the intermediate filament family, is a major component of neuronal cytoskeletons. Neurofilaments are dynamic structures; they contain phosphorylation sites for a large number of protein kinases, including protein kinase A, protein kinase C, cyclin-dependent kinase 5, extracellular signal regulated kinase, glycogen synthase kinase-3, and stress-activated protein kinase gamma. In addition to their role in the control of axon caliber, neurofilaments may affect other cytoskeletal elements, such as microtubules and actin filaments. Changes in neurofilament phosphorylation or metabolism are frequently observed in neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), Parkinson's disease, and Alzheimer's disease.

Background References

1. Li D et al. NEFM DNA methylation correlates with immune infiltration and survival in breast cancer. Clin Epigenetics. 2021 May

2. Sohrabi N et al. Regulatory Role of Insulin on Endogenous L1 ORF1 and NEFM Gene Expression through PI3K Signaling Pathway Specifically in Neuroblastoma Cell Line. Iran J Public Health. 2023 Mar

Sequence Similarity

Belongs to the intermediate filament family.

Post-translational Modification

There are a number of repeats of the tripeptide K-S-P, NFM is phosphorylated on a number of the serines in this motif. It is thought that phosphorylation of NFM results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber.; Phosphorylation seems to play a major role in the functioning of the larger neurofilament polypeptides (NF-M and NF-H), the levels of phosphorylation being altered developmentally and coincidentally with a change in the neurofilament function.; Phosphorylated in the head and rod regions by the PKC kinase PKN1, leading to the inhibition of polymerization.

Subcellular Location

Cell projection, Cytoplasm, Cytoskeleton, Intermediate filament.

UNIPROT

SwissProt: P07197 Human

SwissProt: P08553 Mouse

SwissProt: P12839 Rat

Synonyms

150kDa medium antibody

NEF3 antibody

NEFM antibody

Neurofilament 3 antibody

NF160 antibody

NFM antibody

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 mouse cerebellum tissue with Rabbit anti-NF-M antibody (<a href="/products/HA751854" style="font-weight: bold;text-decoration: underline;">HA751854</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/HA751854" style="font-weight: bold;text-decoration: underline;">HA751854</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 Rabbit anti-NF-M antibody (HA751854) 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 (HA751854) 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 Rabbit anti-NF-M antibody (<a href="/products/HA751854" style="font-weight: bold;text-decoration: underline;">HA751854</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/HA751854" style="font-weight: bold;text-decoration: underline;">HA751854</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 Rabbit anti-NF-M antibody (HA751854) 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 (HA751854) 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 Rabbit anti-NF-M antibody (<a href="/products/HA751854" style="font-weight: bold;text-decoration: underline;">HA751854</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/HA751854" style="font-weight: bold;text-decoration: underline;">HA751854</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 Rabbit anti-NF-M antibody (HA751854) 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 (HA751854) 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-M on different lysates with Rabbit anti-NF-M antibody (<a href="/products/HA751854" style="font-weight: bold;text-decoration: underline;">HA751854</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T cell lysate (20 µg/Lane)<br />Lane 2: Rat brain tissue lysate (40 µg/Lane)<br />Lane 3: Rat liver tissue lysate (negative) (40 µg/Lane)<br />Lane 4: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 5: Mouse liver tissue lysate (negative) (40 µg/Lane)<br /><br />Predicted band size: 102 kDa<br />Observed band size: 150 kDa<br /><br />Exposure time: Lane 1-3: 3 minutes; Lane 4-5: 18 seconds; ECL: K1801;<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/HA751854" style="font-weight: bold;text-decoration: underline;">HA751854</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of NF-M on different lysates with Rabbit anti-NF-M antibody (HA751854) at 1/1,000 dilution.

    Lane 1: 293T cell lysate (20 µg/Lane)
    Lane 2: Rat brain tissue lysate (40 µg/Lane)
    Lane 3: Rat liver tissue lysate (negative) (40 µg/Lane)
    Lane 4: Mouse brain tissue lysate (40 µg/Lane)
    Lane 5: Mouse liver tissue lysate (negative) (40 µg/Lane)

    Predicted band size: 102 kDa
    Observed band size: 150 kDa

    Exposure time: Lane 1-3: 3 minutes; Lane 4-5: 18 seconds; 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 (HA751854) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) 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"