| Source: |
Rat EPO, amino acids (Ala27-Arg192) (Accession# D10763) was expressed in CHO cells. |
| Molecular Mass: |
The 166 amino acid recombinant protein has a predicted molecular mass of approximately 18.5 kD. The DTT-reduced and non-reduced glycosylated protein migrate at approximately 35-50 kD by SDS-PAGE. The N-terminal amino acid is Ala. |
| Purity: |
>95%, as determined by Coomassie stained SDS-PAGE. |
| Formulation: |
0.22 µm filtered protein solution is in 1X PBS, pH 7.2. |
| Endotoxin Level: |
Less than 0.01 ng per µg cytokine as determined by the LAL method. |
| Concentration: |
10 - 100 µg sizes are bottled at 200 µg/mL. 500 µg and larger sizes are bottled at the concentration indicated on the vial. |
| Storage & Handling: |
Unopened vial can be stored between 2°C and 8°C for three months, at -20°C for six months, or at -70°C for one year. For maximum results, quick spin vial prior to opening. Stock solutions should be prepared at no less than 10 µg/mL in sterile buffer (PBS, HPBS, DPBS, and EBSS) containing carrier protein such as 1% BSA or HSA. After dilution, the cytokine can be stored between 2°C and 8°C for one month or from -20°C to -70°C for up to 3 months. Avoid repeated freeze/thaw cycles. |
| Activity: |
ED50 = 0.5 - 2.5 ng/ml, corresponding to a specific activity of 0.4 - 2.0 x 106 units/mg, as determined by induction of TF-1 cell proliferation. |
| Application: |
Bioassay |
|
TF-1 cell proliferation induced by rat EPO.
|
|
| Description: |
Rat Erythropoietin was cloned in 1992, and it has a 79% and 95% homology with human and mouse EPOs, respectively. EPO is a glycoprotein composed of 40-60% carbohydrates, and its molecular weight varies from 30-34 kD depending on the carbohydrate percentage. Hypoxia induces erythropoiesis, and hypoxia inducible factor (HIF) is directly involved in EPO expression. HIF is a heterodimeric transcription factor (HIF-1α, HIF-2α, and beta subunits), and it is regulated by HIF-prolyl hydroxylase (HIF-PH) that hydroxylates the alpha subunits targeting them for ubiquitination and subsequent degradation. HIF-PH is a sensor of levels of iron, oxygen, and metabolic activity. High levels of HIF protein induce EPO production in the kidney and liver, and mobilization of iron to support erythropoiesis. EPO binds to EPOR, and transcripts of this receptor have been detected in non-erythroid precursors such as endothelial cells and renal proximal epithelial cells; nevertheless, it has been published that functional erythropoietin receptor is not detected in endothelial, cardiac, neuronal, and renal cells. Several transcriptions factors play a role in the expression of EPOR such as GATA1, Friend of GATA (Fog1), and the erythroid specific factor SCL/Tal1. EPO has been used to treat anemia associated with chronic kidney disease, cancer patients on chemotherapy, and as an antiviral HIV therapy. |
| Distribution: |
EPO is primarily expressed in the kidney (70-90% of the total EPO) and liver. EPO is expressed by hepatocytes, cortical interstitial fibroblasts in ischemic kidneys, astrocytes, and neurons under hypoxia. |
| Function: |
EPO is the major regulator of differentiation, proliferation, and survival of erythroid progenitors. EPO expression is induced by hypoxia; HIF, among other transcription factors, regulates the EPO expression. EPO has a possible role in neuroprotection. |
| Ligand Receptor: |
EpoR (homodimer). |
| Interaction: |
Erythroid progenitor cells. |
Antigen
References: |
1. Nagao M, et al. 1992. Biochim. Biophys. Acta. 1171:99.
2. Chin K, et al. 2000. Brain Res. Mol. Brain Res. 81:29.
3. Snow JW and Orkin SH. et al. 2009. J. Biol. Chem. 284:29310.
4. Kassouf MT, et al. 2010. Genome Res. 20:1064.
5. Paliege A, et al. 2010. Kidney Int. (Nature) 77:312.
6. Sinclair AM, et al. 2010. Blood 115:4264.
7. Mawadda A, et al. 2012. Anatomy Research International 2012: Article ID 953264. |
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