NENF Human

Neudesin Neurotrophic Factor Human Recombinant
Cat. No.
BT12101
Source
Escherichia Coli.
Synonyms
Neudesin, Cell immortalization-related protein 2, Neuron-derived neurotrophic factor, Secreted protein of unknown function, SPUF protein, NENF, CIR2, SPUF, SCIRP10.
Appearance
Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 95.0% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

NENF Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain (aa 32-172) containing 151 amino acids including a 10 a.a N-terminal His tag and having a molecular mass of 16.9kDa.

Product Specs

Introduction
Neudesin Neurotrophic Factor (NENF), a member of the cytochrome b5 family and specifically the MAPR subfamily, possesses a cytochrome b5 heme-binding domain. This factor demonstrates neurotrophic activity by triggering the phosphorylation of MAPK1/ERK2, MAPK3/ERK1, and AKT1/AKT in primary cultured neurons. However, it lacks mitogenic activity in primary cultured astrocytes. NENF is believed to be involved in neuronal differentiation and may have a transient impact on neural cell proliferation within neural precursor cells. Its neurotrophic activity is enhanced upon binding to heme. Interestingly, NENF expression is elevated in immortal cells and is induced in estrogen receptor-positive breast cancer cells that express the progesterone receptor.
Description

Recombinant human NENF, expressed in E. coli, is a single, non-glycosylated polypeptide chain consisting of 151 amino acids (aa 32-172). It includes a 10-amino acid N-terminal His tag and has a molecular weight of 16.9 kDa.

Physical Appearance
White powder, lyophilized and filtered.
Formulation
The NENF protein underwent filtration (0.4 μm) and subsequent lyophilization from a solution of 0.5 mg/ml in 0.05 M phosphate buffer containing 0.075 M NaCl at pH 7.4.
Solubility

To prepare a working stock solution, it is advised to add deionized water to the lyophilized pellet, aiming for a concentration of approximately 0.5 mg/ml. Allow the pellet to dissolve fully. Note that the NENF product is not sterile. Prior to using it in cell culture, it is essential to filter the solution using an appropriate sterile filter.

Stability
Lyophilized NENF protein should be stored at -20°C. Once reconstituted, aliquot the protein to minimize repeated freeze-thaw cycles. The reconstituted protein exhibits stability at 4°C for a limited period; it remains unchanged for two weeks when stored at this temperature.
Purity
The purity of the protein is greater than 95.0% as determined by SDS-PAGE analysis.
Synonyms
Neudesin, Cell immortalization-related protein 2, Neuron-derived neurotrophic factor, Secreted protein of unknown function, SPUF protein, NENF, CIR2, SPUF, SCIRP10.
Source
Escherichia Coli.
Amino Acid Sequence
MKHHHHHHASGQTPRPAERG PPVRLFTEEE LARYGGEEED QPIYLAVKGV VFDVTSGKEF YGRGAPYNAL TGKDSTRGVA KMSLDPADLT HDTTGLTAKE LEALDEVFTK VYKAKYPIVG YTARRILNED GSPNLDFKPE DQPHFDIKDE F.

Product Science Overview

Structure and Synthesis

Human Neudesin is synthesized as a 172 amino acid precursor, which includes a 31 amino acid signal sequence and a 141 amino acid mature region . The mature region contains a cytochrome b5-like heme-binding domain spanning amino acids 44-129 and a lysine acetylation site at amino acid 136 . The attachment of hemin to its heme-binding domain is essential for its neurotrophic activity .

Molecular Characteristics
  • Molecular Weight: The predicted molecular mass of Neudesin is approximately 16.5 kDa, but it appears as 19-21 kDa under reducing conditions in SDS-PAGE due to its glycosylation .
  • Form: Neudesin is typically found in a monomeric form .
Biological Activity

Neudesin exhibits neurotrophic activity, which means it supports the growth, survival, and differentiation of neurons . This activity is mediated through the binding of heme to its heme-binding domain . Neudesin has been shown to activate the MAPK and PI-3K signaling pathways, which are crucial for cell proliferation and survival .

Recombinant Production

Recombinant human Neudesin is produced using Chinese Hamster Ovary (CHO) cells . The recombinant protein is often tagged with a C-terminal 6-His tag to facilitate purification . The protein is typically lyophilized from a 0.2 μm filtered solution in PBS and can be reconstituted at 500 μg/mL in PBS .

Applications

Recombinant Neudesin is used in various research applications, including:

  • Cell Proliferation Assays: It is measured in cell proliferation assays using Neuro-2A mouse neuroblastoma cells .
  • Neurotrophic Studies: Due to its neurotrophic properties, it is used in studies related to neuronal growth and differentiation .
Storage and Stability

The recombinant protein is stable for 12 months from the date of receipt when stored at -20 to -70°C as supplied . After reconstitution, it remains stable for one month at 2 to 8°C under sterile conditions and for three months at -20 to -70°C under sterile conditions .

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