Certificate of Analysis and Data Sheet

Source:
E.coli

Catalog No.
PRO-359

Background:

Description :

Recombinant Human Collagen produced in E.Coli is a non-glycosylated polypeptide chain a molecular mass of 96,000 Dalton containing human collagen旧 cDNA transcribed reversely from mRNA.
Recombinant Collagen is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation:

The protein was lyophilized after from a sterile solution containing no additives.

Solubility:

It is recommended to reconstitute the lyophilized Human Collagen in sterile 18MΩ-cm H₂O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.

Stability:

Lyophilized rHuCollagen although stable at room temperature for 3 weeks, should be stored desiccated below -18⁰C. Upon reconstitution rHuCollagen should be stored at 4⁰C between 2-7 days and for future use below -18⁰C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid 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 gel.

Amino acid sequence:

The sequence of the first fifteen N-terminal amino acids was determined and was found to be MDPVVLQRRDWENPG.

Amino acid %:

Asp 0.39; Thr 0.16; Ser 5.70; Pro 18.87; Gly 29.17; Ala 13.38; Val .71; Met .18; Ile 0.16; Leu 8.04; Tyr 0.90; Phe 0.15; Lys 0.27; His 0.15; Arg 0.03; Gln 17.40; Hydro Pro 4.34.

Biological Activity:

1. Cultivation of BHK-21 and 2BS cell line BHK-21 or 2BS cells were plated onto the bottom of the 96-well cell culture plates at the density of 1*103cells/well, respectively, with 0.1ml RPMI1640 contained 10% fetal bovine serum in the well. BHK-21 or 2BS cells were incubated in standard conditions (at 37C with 5% CO2 in a humidified incubator) for 24 hours.

2. Biochemical activity analysis of Human-like Collagen) Complete culture media was removed after 24 hours, rHuCollagen was added at various concentrations 0.01%,0.03%,0.05%,0.07%,0.09%,0.10%,0.11%,0.13% and a complete medium was used as blank control. Cells were incubated in standard conditions (at 37C with 5% CO2 in a humidified incubator). The morphology of cell was observed by reverse microscope after 24hr, 48hr, 72hr, 96hr, and 120hr.
Thereafter, the viable cell count was determined by a colorimetric method using MTT (3-4,5dimethylthiazol-2yl-2, 5 diphenyl tetrazolium bromide) after 24hr, 48hr, 72hr, 96hr and 120hr. A490 was evaluated by ELISA reader.
3. Results The results demonstrate that rHuCollagen has no significant effect on cell morphology. MTT analysis shows that rHuCollagen has remarkable effect on cell proliferation when its concentration is higher than 0.03%. Cell proliferation was improved by 40% to 60% compared to control.

Endotoxin:

Less than 0.1 ng/ug (IEU/ug) of Recombinant Human Collagen.

Protein content:

Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm.
2. Analysis by RP-HPLC.

Various uses of activity:

A. Bio-Absorption
B. Cell-Adhesion
C. Mini-immunogenicity and neovascularization
D. Promoting the formation of epithelial cell
E. Biocompatibility
F. Absorption water
G. Against UV radiation.
H. Repairing defected skin
I. Removing pock, acne, spot
J. Dispelling black eye sockets, eye dropping

Usage:

Prospec'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.

Latest Publications:

1. Collagen metabolism in the uterosacral ligaments and vaginal skin of women with uterine prolapse.
BJOG 2006 Jan;113(1):39-46
2. Collagen ultrastructure and TGF-beta1 expression preserved with aminoguanidine during wound healing in diabetic rats.
Endocr Res 2005;31(3):229-43
3. Collagen structure in skin from fibromyalgia patients.
Int J Tissue React 2005;27(3):75-82
4. Treatment of genital prolapse by hammock using porcine skin collagen implant (Pelvicol).
Urology 2005 Dec;66(6):1314-8
5. Molecular mechanisms of junctional epidermolysis bullosa: Col 15 domain mutations decrease the thermal stability of collagen XVII.
J Invest Dermatol 2005 Dec;125(6):1112-8
6. Signalling and regulation of collagen I synthesis by ET-1 and TGF-beta1.
FEBS J 2005 Dec;272(24):6297-309