GDF5 Human
Growth Differentiation Factor-5 Human Recombinant
Cat. No.
BT18292
Source
Escherichia Coli.
Synonyms
Cartilage-derived morphogenetic protein-1, CDMP-1, LAP4, SYNS2, GDF-5, Radotermin, CDMP1, GDF5, Growth differentiation factor 5, BMP-14.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
(a) Analysis by RP-HPLC.
(b) Analysis 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
Growth Differentiation Factor 5 Human Recombinant produced in E.Coli is a homodimer, non-glycosylated polypeptide chain containing 2 x 120 amino acids and having a total molecular mass of 27.4kDa. To enable bacterial expression the N-terminal sequence of Ala-Pro-Leu-Thr was replaced with a Lys.
GDF5 is purified by proprietary chromatographic techniques.
GDF5 is purified by proprietary chromatographic techniques.
Product Specs
Introduction
Growth Differentiation Factor 5 (GDF-5), a protein belonging to the bone morphogenetic protein (BMP) family within the TGF-beta superfamily, plays a crucial role in regulating cell growth and differentiation in both embryonic and adult tissues. GDF-5 is synthesized as a precursor protein that undergoes proteolytic processing to release a mature protein containing seven conserved cysteine residues. This mature protein is involved in skeletal development, as evidenced by the association of GDF-5 gene mutations with skeletal dysplasias like acromesomelic dysplasia, Hunter-Thompson type; brachydactyly, type C; and chondrodysplasia, Grebe type.
Description
Recombinant Human Growth Differentiation Factor 5, expressed in E. coli, is a non-glycosylated homodimeric protein. Each subunit comprises 120 amino acids, resulting in a total molecular mass of 27.4 kDa. To facilitate bacterial expression, the N-terminal sequence has been modified by replacing the original Ala-Pro-Leu-Thr sequence with Lys. The protein undergoes purification using proprietary chromatographic techniques.
Physical Appearance
White, lyophilized powder, sterile-filtered.
Formulation
The lyophilized Growth Differentiation Factor 5 is supplied without any additional additives.
Solubility
To reconstitute the lyophilized Growth Differentiation Factor-5, it is recommended to dissolve it in sterile 18 MΩ-cm H2O at a concentration of at least 100 µg/ml. This solution can then be further diluted in other aqueous solutions as needed.
Stability
Lyophilized Growth Differentiation Factor 5 remains stable at room temperature for up to 3 weeks; however, it is recommended to store it desiccated at a temperature below -18°C. After reconstitution, Growth Differentiation Factor-5 should be stored at 4°C for a period of 2-7 days. For long-term storage, it is advisable to store it below -18°C. The addition of a carrier protein such as HSA or BSA (0.1%) is recommended for long-term storage. It is important to avoid repeated freeze-thaw cycles.
Purity
The purity is determined to be greater than 98.0% through the following methods:
(a) Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) analysis.
(b) Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis.
(a) Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) analysis.
(b) Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis.
Biological Activity
The biological activity of GDF-5 is determined by its ability to induce alkaline phosphatase activity in ATDC5 cells. Typically, the activity is observed at concentrations of 10-20 ng/ml.
Protein Content
Protein quantification was performed using two independent methods:
1. UV spectroscopy at 280 nm, utilizing an absorbance value of 1.15 as the extinction coefficient for a 0.1% (1 mg/ml) solution. This value was determined using the PC GENE computer analysis program (IntelliGenetics) for protein sequence analysis.
2. Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) analysis, employing a standard solution of GDF5 as a reference standard.
1. UV spectroscopy at 280 nm, utilizing an absorbance value of 1.15 as the extinction coefficient for a 0.1% (1 mg/ml) solution. This value was determined using the PC GENE computer analysis program (IntelliGenetics) for protein sequence analysis.
2. Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) analysis, employing a standard solution of GDF5 as a reference standard.
Synonyms
Cartilage-derived morphogenetic protein-1, CDMP-1, LAP4, SYNS2, GDF-5, Radotermin, CDMP1, GDF5, Growth differentiation factor 5, BMP-14.
Source
Escherichia Coli.
Amino Acid Sequence
APSATRQGKR PSKNLKARCS RKALHVNFKD MGWDDWIIAP LEYEAFHCEG LCEFPLRSHL EPTNHAVIQT LMNSMDPEST PPTCCVPTRL SPISILFIDS ANNVVYKQYE DMVVESCGCR.
Product Science Overview
Structure and Function
GDF-5 is synthesized as a precursor protein that undergoes proteolytic cleavage to produce a mature protein containing seven conserved cysteine residues . The mature GDF-5 protein forms a disulfide-linked homodimer, which is essential for its biological activity . The molecular weight of the recombinant human GDF-5 protein is approximately 27.4 kDa .
Biological Role
GDF-5 plays a critical role in the development of bones, joints, and limbs . It binds to a receptor complex comprising BMPR1B and BMPR2, which then activates the SMAD signaling pathway . This signaling pathway is crucial for the regulation of gene expression involved in bone and cartilage formation.
Clinical Significance
Alterations in the expression of GDF-5 have been linked to various skeletal disorders. For instance, decreased or altered expression of GDF-5 is associated with the pathogenesis of osteoarthritis and certain forms of dwarfism . Additionally, GDF-5 has been implicated in the development of Parkinson’s disease .
Recombinant GDF-5
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