Anti-NFM (Neurofilament Medium Chain) Antibody

NFM protein was purified from bovine spinal cords following the method of Delacourte et al. ["Study of the 10-nm-filament fraction isolated during the standard microtubule prepartion" (1980) . Biochem. J. 191 (2) : 543-546], followed by Prepcell purification (Bio-Rad) . Antibodies were prepared by injecting purified NFM protein into laying hens and purifying the IgY fraction from eggs collected from hyper-immunized hens. After several boosts, hens were co-boosted with recombinant NFM protein expressed in bacteria. These antibodies have been validated with human, mouse and rat tissues.

Product Specifications

Background

Human Neurofilament Medium Chain (NFM) is a 102,448 dalton protein (916 amino acids) expressed in CNS and PNS neurons. This cytoskeletal protein is normally phosphorylated in vivo and its extent of phosphorylation correlates with different states of axonal growth and stabilization. These observations have led to the hypothesis that the NFM protein cross-links adjacent neurofilament strands and contributes to the structural integrity of the axon.

Synonyms

Neurofilament medium polypeptide, NF-M, 160 kDa neurofilament protein, Neurofilament 3, Neurofilament triplet M protein

Other Statements

These antibodies are to be used as research laboratory reagents and are not for use as diagnostic or therapeutic reagents in humans.

Gene Name

NEFM NEF3 NFM

UniProt

P07197

Host

Chicken

Species Reactivity

Human, Mouse, Rat

Reactivity

Human, Mouse, and Rat

Immunogen

Recombinant construct containing the C-terminus of the human sequence expressed in Escherichia coli

Immunogen Type

Human

Target

Neurofilament Medium Chain (NFM)

Clonality

Polyclonal

Isotype

IgY

Conjugation

Unconjugated

Applications

IHC, ICC, WB

Field of Research

Cell Markers Neuronal Markers Axonal Markers

Assay Type

Primary Antibody

Assay Protocol

Chickens were immunized with NFM protein was purified from bovine spinal cords. After repeated injections into the hens, immune eggs were collected, and the IgY fractions were purified from the yolks.

Concentration

2 mg/mL

Format

IgY Fraction

Form

IgY Fraction

Buffer

Phosphate-buffered (10 mM) isotonic (0.9%, w/v) saline ("PBS" pH 7.2) with bovine serum albumin (BSA, 0.5%) added to prevent absorption to the plastic and sodium azide (0.02%, w/v) added as a preservative

Molecular Weight

150 kDa

Precautions

These antibodies are to be used as research laboratory reagents and are not for use as diagnostic or therapeutic reagents in humans.

Shipping Conditions

Ice Packs

Storage Conditions

Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least twelve months, provided they remain sterile. For longer term storage, aliquot and freeze to avoid freeze-thaw of the antibody.

Shelf Life

12 months from receipt of product

IHC Dilution

1:1000-1:2000

WB Dilution

1:1000-1:2000

Pre Treatment

Target Description

ICC Dilution

1:1000-1:2000

Quality Control

Quality assurance analysis was performed using immunohistochemistry (at a dilution of 1:5000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.