Description:
Recombinant Human GRO-alpha produced in E.Coli is a single,non-glycosylated, polypeptide chain containing 73 amino acids and having a molecular mass of 7811 Dalton. rHuGRO-alpha is purified by proprietary chromatographic techniques.

Source:
Escherichia Coli.

Formulation:
The protein was lyophilized with no additives.

Solubility:
It is recommended to reconstitute the lyophilized rHuGRO-alpha in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.

Stability:
Lyophilized rHuGRO-? although stable at room temperature for 3 weeks, should be stored desiccated below -18C. Upon reconstitution rHuGRO-alpha should be stored at 4C between 2-7 days and for future use below -18C.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.

Purity:
Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained.

Amino acid sequence:
The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Ser-Val-Ala-Thr.

Dimers and aggregates:
Less than 1% as determined by silver stained SDS-PAGE gel analysis.

Biological Activity:
Recombinant Melanoma growth stimulatory activity is fully biologically active when compared to standard. The biological activity was determined by measuring the dose dependent mobilization of intracellular calcium (calcium flux) with human neutrophils. Significant calcium mobilization is observed with ≥1ng/mL of recombinant human GRO-alpha. The optimal concentration for each specific application should be determined by an initial dose-response assay.

Endotoxin:
Less than 0.1 ng/µg (IEU/µg) of rHuGRO-alpha.

Protein content:
Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm.
2. Analysis by RP-HPLC, using a standard solution of GRO-alpha as a Reference Standard.