Recombinant Human Adiponectin/Acrp30 Protein Summary
| Description |
A biologically active protein corresponding to the signal peptide sequence, and the mature Adiponectin sequence of ADIPOQ.Source: modified human 293 cells |
| Specificity |
A DNA sequence encoding the human Adiponectin protein sequence. Adiponectinmigrates as a broad band between 25 and 35 kDa in SDS-PAGE due to post-translational modifications, in particular glycosylation. This compares with unmodified Adiponectin that has a predicted molecular mass of 24.5 kDa. Adiponectin contains O-linked oligosaccharides. |
Preparation Method |
A DNA sequence encoding the human Adiponectin protein sequence (containing the signal
peptide sequence, and the mature Adiponectin sequence) was expressed in modified
human 293 cells. |
| Details of Functionality |
The ED50 of adiponectin is typically 3-5 ug/ml as measured by its ability to inhibit M1
cell proliferation. |
| Protein/Peptide Type |
Recombinant Protein |
| Gene |
ADIPOQ |
Applications/Dilutions
| Dilutions |
- Bioactivity
- Block/Neutralize
- Functional
- Western Blot
|
| Application Notes |
Adiponectin separates into a number of glycoforms with an observed pI between 4.5 and 8.0 on 2D PAGE due to post-translational modifications, in particular glycosylation. This compares with the unmodified Adiponectin that has a predicted pI of 5.5. |
Reactivity Notes
Packaging, Storage & Formulations
| Storage |
Store at -80C. Avoid freeze-thaw cycles. |
| Concentration |
LYOPH |
| Reconstitution Instructions |
Reconstitute with 0.5 ml sterilized PBS. After reconstitution the buffer solution will contain 1% human serum albumin and 10% trehalose already present in the vial. |
Alternate Names for Recombinant Human Adiponectin/Acrp30 Protein
Background
Adiponectin is a member of the 'adipocytokines' that are cytokines that are expressed from adipose tissues. Its general functions include regulation of energy homeostasis, hematopoiesis, inflammation and immunity. Adiponectin also plays an important role in fat metabolism, feeding behaviour, insulin sensitivity and is a negative regulator of hematopoiesis and immune responses. Adiponectin has been shown to suppress the expression of a number of membrane bound proteins involved in the infiltration of cells to sites of inflammation, including, vascular cell adhesion molecule 1 (VCAM-1), E-Selectin, and intercellular adhesion molecule-1 (ICAM-1). Furthermore adiponectin suppresses TNF induced adhesion of monocytes to endothelial cells through a mechanism involving the suppression of TNF-alpha induced I-kappa-B-alpha phosphorylation. Adiponectin also inhibits phagocytosis and bacterial lipopolysaccharideinduced production of TNF-alpha in mature macrophages. Conversely, TNF-alpha reduces the expression and secretion of adiponectin in differentiating primary human pre-adipocytes. This is a HCX protein. HCX Expression System Details HCX proteins mimic the proteins in the human body because they are expressed from human, rather than animal, insect or bacterial cells. This process gives them human post-translational modifications. Recombinant DNA techniques allow a human protein with the correct amino acid sequence to be expressed in a non-human cell line. However, non-human cells lack the appropriate cellular machinery, such as specific glycosyltransferases, necessary to produce the correct human post-translational modifications of a protein. An extreme example is seen in E. coli cells, which produce recombinant proteins with no glycosylation, as the above figure illustrates. Rodent and yeast cells are able to glycosylate proteins, but they are still different from glycosylation in human cells. Expression System Resultant Proteins Human (e.g. K562, HEK293) Correct amino acid sequence Human post-translational modifications Rodent (e.g. CHO, NSO) Correct amino acid sequence Some natural glycosylation - not human-like Yeast (e.g. Pichia) Correct amino acid sequence Some natural glycosylation - not human-like E.Coli Correct amino acid sequence No PTMs Although there have been significant attempts to make non-human cell derived cytokines more human-like, there is a growing awareness that in many instances, particularly in therapeutics, cytokines should mimic those found in the body as closely as possible. Purified Adiponectin consists of 0 to 30% carbohydrate by weight.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Peptides and proteins are
guaranteed for 1 year from date of receipt.
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I'd like to confirm that the recommended ND50 is 5 micrograms per millilitre-is this correct?
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