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- Jan 25, 2014
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Evaluation of recombinant human IGF-1 activity through the conversion of resazurin to resofurin by the Neuro2a mouse cell line.
Resazurin is an aromatic compound, a redox indicator - a blue dye, which, upon reduction, turns into a pink fluorescent resorufin. Cells, including the Neuro2a line, reduce resazurin during metabolism. The more active the metabolism is, the more active is the reduction to fluorescent resorufin. Thus, the intensity of the cells glowing in ultraviolet light is directly proportional to the activity of the added hormone.
The plate with cells in serum-free medium was supplemented with
• Recombinant IGF-1 (Insubolic) at 100 ng / ml concentration;
• Pharmacopoeia standard Sigma at 100 ng / ml concentration;
• Blood serum at 10% concentration.
After incubation, resazurin was added to the cells and incubated for conversion (4 hours). The luminescence was assessed visually (B) and using a BMG Clariostar fluorimeter according to the standard "Resarufin" protocol (A). Also, the state and size of the cells were assessed microscopically (C).
According to the obtained data, the Insubolic IGF-1 demonstrated high activity (higher than in the sample with added serum), and slightly higher than the standard from Sigma. The micrographs show an increase in the volume of cells, as well as a change in structure (morphological changes characteristic of neuronal cells occur).
Gents, sharing this very interesting lab test.
There're 4 tubes :
1 Tube : nothing was added.
2 Tube blood serum at 10% concentration
3 The Human Recombinant IGF-1 obtained from Sigma Aldrich (known and respected supplier of lab material) at 100 ng/ml concentration
4 Insubolic IGF-1 (Qore Pharm), 100 ng/ml concentration.
The intensity of the cells glowing in ultraviolet light is directly proportional to the activity of the added hormone.
According to the obtained data, the Insubolic IGF-1 demonstrated high activity (higher than in the sample with added serum), and slightly higher than the standard from Sigma. The micrographs show an increase in the volume of cells, as well as a change in structure (morphological changes characteristic of neuronal cells occur).
Basically, Insubolic IGF-1 slightly outperforms the Sigma Aldrich's IGF-1 in terms of it's bioactivity.
This also proves that Qore Pharm has a team capable of creating formulations that are on par or better than some known big pharma brands.
I'm really proud of the results! =)
RS
Resazurin is an aromatic compound, a redox indicator - a blue dye, which, upon reduction, turns into a pink fluorescent resorufin. Cells, including the Neuro2a line, reduce resazurin during metabolism. The more active the metabolism is, the more active is the reduction to fluorescent resorufin. Thus, the intensity of the cells glowing in ultraviolet light is directly proportional to the activity of the added hormone.
The plate with cells in serum-free medium was supplemented with
• Recombinant IGF-1 (Insubolic) at 100 ng / ml concentration;
• Pharmacopoeia standard Sigma at 100 ng / ml concentration;
• Blood serum at 10% concentration.
After incubation, resazurin was added to the cells and incubated for conversion (4 hours). The luminescence was assessed visually (B) and using a BMG Clariostar fluorimeter according to the standard "Resarufin" protocol (A). Also, the state and size of the cells were assessed microscopically (C).
According to the obtained data, the Insubolic IGF-1 demonstrated high activity (higher than in the sample with added serum), and slightly higher than the standard from Sigma. The micrographs show an increase in the volume of cells, as well as a change in structure (morphological changes characteristic of neuronal cells occur).
Gents, sharing this very interesting lab test.
There're 4 tubes :
1 Tube : nothing was added.
2 Tube blood serum at 10% concentration
3 The Human Recombinant IGF-1 obtained from Sigma Aldrich (known and respected supplier of lab material) at 100 ng/ml concentration
4 Insubolic IGF-1 (Qore Pharm), 100 ng/ml concentration.
The intensity of the cells glowing in ultraviolet light is directly proportional to the activity of the added hormone.
According to the obtained data, the Insubolic IGF-1 demonstrated high activity (higher than in the sample with added serum), and slightly higher than the standard from Sigma. The micrographs show an increase in the volume of cells, as well as a change in structure (morphological changes characteristic of neuronal cells occur).
Basically, Insubolic IGF-1 slightly outperforms the Sigma Aldrich's IGF-1 in terms of it's bioactivity.
This also proves that Qore Pharm has a team capable of creating formulations that are on par or better than some known big pharma brands.
I'm really proud of the results! =)
RS
