pGH 22kDa Human
Growth Hormone Placental 22kDa Human Recombinant
Cat. No.
BT15319
Source
Escherichia Coli.
Synonyms
GHL, GHV, GH-V, hGH-V, PGH.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 98.0% as determined by:
(a) Analysis by SEC-HPLC.
(b) Analysis by SDS-PAGE.
(a) Analysis by SEC-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
Placental HGH 22kDa Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 192 amino acids and having a molecular mass of 22367 Dalton. Predicted pI=7.80. Placental Growth Hormone has dimished lactogenic (prolactin receptor mediated) activity characteristic to pituitary GHs.
GH Placental Human Recombinant is purified by proprietary chromatographic techniques.
GH Placental Human Recombinant is purified by proprietary chromatographic techniques.
Product Specs
Introduction
Growth hormone (GH) belongs to the somatotropin/prolactin family of hormones and plays a crucial role in regulating growth. The GH gene, along with four other related genes, is found on chromosome 17 at the growth hormone locus. These genes share the same transcriptional orientation, suggesting their evolution through gene duplication events. Notably, they exhibit a high degree of sequence similarity. Alternative splicing further diversifies these five growth hormones, generating various isoforms with potential functional specializations. This particular family member is expressed in the pituitary gland but, unlike the other four genes in the growth hormone locus, not in placental tissue. Mutations or deletions in the GH gene can lead to growth hormone deficiency, resulting in short stature.
Description
Recombinant Placental HGH 22kDa, produced in E. coli, is a single, non-glycosylated polypeptide chain. It comprises 192 amino acids, resulting in a molecular mass of 22,367 Daltons. The predicted isoelectric point (pI) is 7.80. Unlike pituitary GHs, Placental Growth Hormone exhibits diminished lactogenic activity (mediated by the prolactin receptor). The purification process of Recombinant Human Placental GH involves proprietary chromatographic techniques.
Physical Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation
The protein was lyophilized from a concentrated solution (1 mg/mL) containing 0.0045 mM NaHCO3, with the pH previously adjusted to 8-9.
Solubility
To reconstitute the lyophilized Placental HGH, it is recommended to dissolve it in 0.4% NaHCO3 or water adjusted to pH 9. The initial reconstitution concentration should be at least 100 µg/mL. This solution can then be further diluted into other aqueous solutions as needed.
Stability
Lyophilized placental GH remains stable at room temperature for up to 3 weeks. However, for long-term storage, it is recommended to store it desiccated at a temperature below -18°C. Once reconstituted and sterilized by filtration, placental HGH can be stored at 4°C for up to 4 weeks. For extended storage periods or when working with more diluted solutions, adding a carrier protein (0.1% HSA or BSA) is advised. Avoid repeated freeze-thaw cycles.
Purity
The purity of this product is greater than 98.0%, as determined by the following methods:
(a) Size Exclusion Chromatography - High Performance Liquid Chromatography (SEC-HPLC) analysis.
(b) Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis.
(a) Size Exclusion Chromatography - High Performance Liquid Chromatography (SEC-HPLC) analysis.
(b) Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis.
Protein Content
Protein quantitation was performed using two independent methods:
1. UV spectroscopy at 280 nm, employing an extinction coefficient of 1.18 for a 0.1% (1 mg/mL) solution. This value was calculated using the PC GENE computer analysis program for protein sequences (IntelliGenetics).
2. Reverse Phase - High Performance Liquid Chromatography (RP-HPLC) analysis, using a calibrated solution of GH-22K-placental as a reference standard.
1. UV spectroscopy at 280 nm, employing an extinction coefficient of 1.18 for a 0.1% (1 mg/mL) solution. This value was calculated using the PC GENE computer analysis program for protein sequences (IntelliGenetics).
2. Reverse Phase - High Performance Liquid Chromatography (RP-HPLC) analysis, using a calibrated solution of GH-22K-placental as a reference standard.
Synonyms
GHL, GHV, GH-V, hGH-V, PGH.
Source
Escherichia Coli.
Amino Acid Sequence
The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Phe-Pro-Thr-Ile.
Product Science Overview
Structure and Production
Synthetic Routes and Reaction Conditions
The production of recombinant placental growth hormone involves several steps:
- Gene Cloning: The gene encoding the placental growth hormone is cloned into an expression vector suitable for E. coli.
- Transformation: The recombinant plasmid is introduced into E. coli cells through a process called transformation.
- Expression: The transformed E. coli cells are cultured under conditions that induce the expression of the recombinant hormone.
- Purification: The recombinant hormone is purified using proprietary chromatographic techniques to achieve a purity greater than 98% .
The lyophilized (freeze-dried) form of the hormone is typically reconstituted in a solution of 0.4% sodium bicarbonate (NaHCO3) or water adjusted to pH 9 . This solution can then be further diluted to other aqueous solutions as needed.
Industrial Production Methods
The industrial production of recombinant placental growth hormone involves large-scale fermentation of E. coli cells containing the recombinant plasmid. The cells are grown in bioreactors under controlled conditions to maximize the yield of the hormone. After fermentation, the cells are harvested, and the recombinant hormone is extracted and purified using chromatographic techniques .
Analysis of Chemical Reactions
The analysis of recombinant placental growth hormone involves several techniques to ensure its purity and activity:
- Size-Exclusion Chromatography (SEC-HPLC): This technique is used to determine the molecular size and purity of the hormone .
- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE): This method is used to analyze the protein’s molecular weight and purity .
- Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC): This technique is used for protein quantitation and to ensure the consistency of the recombinant hormone .
Additionally, the hormone’s biological activity is assessed through various bioassays to confirm its functionality.
Applications and Stability
Recombinant placental growth hormone is used in research to study its role in growth control and its potential therapeutic applications. The lyophilized form of the hormone is stable at room temperature for up to three weeks but should be stored desiccated below -18°C for long-term storage . Upon reconstitution and filter sterilization, the hormone can be stored at 4°C for up to four weeks .
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