Human Erythropoietin-alpha Recombinant
Categories: HematopoietinsRecombinant Human Cytokines$70.00 – $1,450.00
Description
Accession
P01588
Source
Optimized DNA sequence encoding Human EPO mature chain was expressed in CHO cells.
Molecular weight
Native human Erythropoietin-alpha is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 18kDa. Recombinant EPO-a is a glycosylated monomer protein consisting of 166 amino acid residue subunits, and due to glycosylation migrates as an approximately 35 kDa protein under reducing conditions in SDS-PAGE.
Purity
>98%, as determined by SDS-PAGE and HPLC
Biological Activity
Activity was determined by the dose-dependent proliferation assay using a factor-dependent human erythroleukemic cell line TF-1 and was found to be.8x105 IU/mg.
Protein Sequence
MGVHECPAWL WLLLSLLSLP LGLPVLGAPP RLICDSRVLE RYLLEAKEAE NITTGCAEHC SLNENITVPD TKVNFYAWKR MEVGQQAVEV WQGLALLSEA VLRGQALLVN SSQPWEPLQL HVDKAVSGLR SLTTLLRALG AQKEAISPPD AASAAPLRTI TADTFRKLFR VYSNFLRGKL KLYTGEACRT GDR
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation
Recombinant Erythropoietin alpha was lyophilized from a 0.2 μm filtered PBS solution pH.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers
Storage
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at2° -8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Interactor
Biological Process
Molecular function
Methods
Effects of overexpression of Bmi1 on HSCs in vitro.
- Single CD34-LSK cells were sorted into 96-well microtiter plates containing the SF-O3 medium supplemented with 10% FBS and multiple cytokines (10 ng/ml SCF, 10 ng/ml TPO, 10 ng/ml IL-3, and 3 u/ml EPO) and allowed to form colonies.
Colony-forming assay
- The effects of SNS-032, perifosine, or combination on the leukemia colony formation (CFU-L) in methylcellulose medium were examined using leukemic colony assay as previously described [3) in 600 μL of methylcellulose solution were incubated in the presence of the agents or an equivalent amount of medium at 37°C in a humidified atmosphere with 5% CO2.
- Primary leukemic cells were cultured in methylcellulose medium containing recombinant human (rh) stem cell factor (SCF), granulocyte macrophage-colony-stimulating factor (GM-CSF), and interleukin 3 (IL-3) at 2 × 104 cells/dish.
- After 7 days, CFU-Ls that contain >40 cells were scored manually under a light microscope .
- For colony assay of human normal bone marrow cells, 3 U/mL rh erythropoietin , 50 ng/mL rhSCF, 30 ng/mL rhGM-CSF, and 10 ng/mL rhIL-3 were added to the methylcellulose medium.
- The colonies were counted under a microscope on day 12 of culture.
Colony forming analysis
- celltype'>Leukemic cell lines, MNCs from the patients with, or healthy controls were infected with or without the indicated viruses (50 MOI) and seeded in triplicate in a mixture containing 1.35% methylcellulose in medium'>Iscove's medium'>modified medium'>Dulbecco's medium (IMDM;) supplemented with 20% fetal bovine serum and10−4 M 2-Mercaptoethanol .
- For colony assay of bone marrow cells, 3 U/mL recombinant human (rh) erythropoietin, 50 ng/mL rh stem cell factor, 30 ng/mL rh granulocyte macrophage–colony-stimulating factor, and 10 ng/mL rh interleukin-3 were added to the methylcellulose medium.
- The colonies were evaluated under a microscope on day 12 of culture.
- In contrast, leukemic cell lines were seeded in cytokine-free mixture as described previously [
