Recombinant Human GDNF GMP Protein, CF

The bioactivity of Recombinant Human GDNF GMP Protein (Catalog # BT-GDNF-GMP) was measured in a cell proliferation assay using the cell proliferation of SH-SY5Y cells. Three independent lots weretested for bioactivity and plotted on the same graph to show lot-to-lot consistency of GMP GDNF protein.

Equivalent bioactivity of GMP (Catalog # BT-GDNF-GMP) and Animal-Free (BT-GDNF-AFL) grades of Recombinant HumanGDNF as measured in a cell proliferation assay using the cell proliferation of SH-SY5Y cells (orange and green, respectively).

2 μg/lane of Recombinant Human GDNF GMP Protein (Catalog # BT-GDNF-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band at 8 kDa under reducing conditions.

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

Recombinant Human GDNF GMP Protein, CF Summary

• Details of Functionality: Measured in a cell proliferation assay using SH‑SY5Y human neuroblastoma cells. The ED₅₀ for this effect is 0.750-9.00 ng/mL in the presence of Recombinant Human GFR alpha -1/GDNF R alpha 1 Fc Chimera (Catalog # 714-GR).
  The specific activity of recombinant human GDNF is >5.0 x 10⁵ units/mg, which is calibrated against the human GDNF Reference Standard (NIBSC code: 09/266).
• Source: E. coli-derived human GDNF protein
  Gly113 - Ile211 with & without an N-terminal Met
  Produced using non-animal reagents in an animal-free laboratory.
  Manufactured and tested under cGMP guidelines.
• N-terminal Sequence: Gly113-Lys-Asn-Arg-Gly-(Cys)-Val-Leu-Thr-Ala & Met-Gly113-Lys-Asn-Arg-Gly-(Cys)-Val-Leu-Thr
• Structure / Form: Disulfide-linked homodimer
• 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 22225 Da ± 5 Da, N-terminal Met species are also present (22356 ± 5 Da)
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 11.3 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: 8 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 22225 Da ± 5 Da, N-terminal Met species are also present (22356 ± 5 Da)
• Reconstitution Instructions: Reconstitute at 500 μg/mL in sterile water.

Notes

Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human GDNF GMP Protein, CF

• Astrocyte-derived trophic factor
• ATF
• ATF1
• ATF2
• GDNF
• glial cell derived neurotrophic factor
• glial cell line derived neurotrophic factor
• glial cell line-derived neurotrophic factor
• HFB1-GDNF
• HGDNF
• HSCR3

Background

Glial Cell Line-derived Neurotrophic Factor (GDNF) is a neurotrophic factor that has been shown to promote the survival of various neuronal subpopulations in both the central as well as the peripheral nervous systems at different stages of their development. Neuronal subpopulations that have been shown to be affected by GDNF include motoneurons, midbrain dopaminergic neurons, Purkinje cells and sympathetic neurons.

Native GDNF, a disulfide-linked homodimeric glycoprotein, is a novel member of the TGF-beta  superfamily. Human GDNF cDNA encodes a 211 amino acid residue prepropeptide that is processed to yield a dimeric protein. Mature human GDNF was predicted to contain two 134 amino acid residue subunits. NS0 expressed mature human GDNF lacks 31 residues from the amino-terminus of the predicted sequence. This glycosylated recombinant mature human GDNF still contains the seven conserved Cys residues found in all members of the TGF-beta superfamily and is biologically active. The GDNF sequence contains two potential glycosylation sites and insect cell‑expressed recombinant rat GDNF proteins are glycosylated. Mature rat and human GDNF exhibit approximately 93% amino acid sequence identity and show considerable species cross-reactivity. Cells known to express GDNF include Sertoli cells, type 1 astrocytes, Schwann cells, neurons, pinealocytes and skeletal muscle cells.