Recombinant Human BDNF GMP Protein, CF

GMP-grade Recombinant Human BDNF Protein (Catalog # BT-BDNF-GMP) as measured in a cell proliferationassay using BaF mouse pro-B cells transfected with TrkB. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lotconsistency of GMP BDNF.

Equivalent bioactivity of GMP (Catalog # BT-BDNFGMP) and Animal-Free (BT-BDNF-AFL) grades of Recombinant Human BDNF asmeasured in a cell proliferation assay using BaF mouse pro-B cells transfected with TrkB (orange and green, respectively).

2 μg/lane of Recombinant Human BDNF GMP Protein (Catalog # BT-BDNF-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 12 kDa, under reducing conditions.

Dopaminergic neurons were successfully differentiated from human iPSCs using Recombinant Human GMP BDNF (Catalog # BT-BDNF-GMP). Neurons show bright co-staining of beta tubulin 3 (TUJ1; green), tyrosine hydroxylase (TH; red), and DAPI (blue) to confirm dopaminergic neuron identity. BDNF successfully supported the derivation and maintenance of dopaminergic neurons.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human BDNF GMP Protein, CF Summary

• Details of Functionality: Measured in a cell proliferation assay using BaF mouse pro-B cells transfected with TrkB. The ED₅₀ for this effect is 0.100-1.00 ng/mL.
  The specific activity of Recombinant Human BDNF is >8.0 x 10⁵ units/mg, which is calibrated against the human BDNF WHO Standard (NIBSC code: 96/534).
• Source: E. coli-derived human BDNF protein
  Arg134 - Arg247, with an N-terminal Met
  Produced using non-animal reagents in an animal-free laboratory.
  Manufactured and tested under cGMP guidelines.
• Accession #: P23560.1
• N-terminal Sequence: Met-Arg134-Arg-Gly-Glu-Leu-Ser-Val-(Cys)-Asp
• Protein/Peptide Type: GMP Recombinant Proteins
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 13121 Da ± 50 Da
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 13.1 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 12 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 13121 Da ± 50 Da
• Reconstitution Instructions: Reconstitute the 25 μg size at 250 μg/mL in PBS. Reconstitute all other sizes at 500 μg/mL in PBS.

Notes

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Alternate Names for Recombinant Human BDNF GMP Protein, CF

• Abrineurin
• ANON2
• BDNF
• brain-derived neurotrophic factor
• BULN2
• MGC34632
• Neurotrophin

Background

Brain-derived neurotrophic factor (BDNF) is a member of the neurotrophin family of growth factors that are required for the differentiation and survival of specific neuronal subpopulations in both the central as well as the peripheral nervous system.

BDNF is often utilized as a cell culture supplement to promote neural differentiation in regenerative medicine studies and clinical manufacturing protocols.

The neurotrophin family is comprised of at least four proteins including BDNF, nerve growth factor (NGF), neurotrophin-3 (NT3), and NT4/5. Human BDNF is initially expressed as a proprotein, which is then cleaved to yield a mature protein. Mature BDNF is a non-covalently linked homodimer, with each monomer containing antiparallel beta -strands and a characteristic cystine knot motif. Within the mature domain, human BDNF shares the identical amino acid sequence with mature mouse and rat BDNF.

BDNF is strongly expressed in various regions of the brain, including the hippocampus and cerebellum, and weaker expression has been detected in the thymus, liver, spleen, heart, and lung. BDNF participates in axonal growth and pathfinding and in the modulation of dendritic growth and morphology and in later stages of development regulates synaptic transmission and plasticity and acts as a central modulator of pain. BDNF binds with high affinity and specifically activates the TrkB tyrosine kinase receptor. BDNF signaling via TrkB is essential for adult synaptic plasticity and the formation of memories.

The BDNF signaling pathway utilizes both AKT and ERK pathways to exert its pleiotrophic effects in the central nervous system. Decreased expression of BDNF is seen in many neurological diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and autism. Further, BDNF is proposed as a biomarker for psychiatric disorders such as bipolar disease and depression and has been implicated in posttraumatic stress disorder, phobia, and panic disorder. A single amino acid substitution, Val66Met, has been shown to lead to reduced, activity-dependent BDNF secretion and memory impairment.

Bioinformatics

• Uniprot:
  • P23560.1()