|The EPO epo (Catalog #MBS2009132) is a Recombinant Protein produced from E Coli and is intended for research purposes only. The product is available for immediate purchase. MyBioSource\'s Erythropoietin (EPO) can be used in a range of immunoassay formats including, but not limited to, SDS-PAGE, Western Blot (WB), ELISA (EIA), Immunoprecipitation (IP). Researchers should empirically determine the suitability of the EPO epo for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process.
The EPO epo product has the following accession number(s) (GI #55742715) (NCBI Accession #NP_999299.1) (Uniprot Accession #P49157). Researchers may be interested in using Bioinformatics databases such as those available at The National Center for Biotechnology Information (NCBI) website for more information about accession numbers and the proteins they represent. Even researchers unfamiliar with bioinformatics databases will find the NCBI databases to be quite user friendly and useful. The amino acid sequence is listed below:
The target protein is fused with two N-terminal Tags, His-tag and GST-tag, its sequence is listed below.
MSPILGYWKI KGLVQPTRLL LEYLEEKYEE HLYERDEGDK WRNKKFELGL EFPNLPYYID GDVKLTQSMA IIRYIADKHN MLGGCPKERA EISMLEGAVL DIRYGVSRIA YSKDFETLKV DFLSKLPEML KMFEDRLCHK TYLNGDHVTH PDFMLYDALD VVLYMDPMCL DAFPKLVCFK KRIEAIPQID KYLKSSKYIA WPLQGWQATF GGGDHPPKSD GSTSGSGHHH HHHSAGLVPR GSTAIGMKET AAAKFERQHM DSPDLGTGGG SGIEGRGSMG YRGSEF-APP R LICDSRVLER YILEAKEGEN ATMGCAESCS FSENITVPDT KVNFYAWKRM EVQQQAMEVW QGLALLSEAI LQGQALLANS SQPSEALQLH VDKAVSGLRS LTSLLRALGA QKEAIPLPDA SPSSATPLRT FAVDTLCKLF RNYSNFLRGK LTLYTGEACR RRDR.
To buy or view more detailed product information and pricing, please click on the technical datasheet page below:
Please refer to the product datasheet for known applications of a given recombinant protein. We\'ve tested the Recombinant Erythropoietin (EPO) with the following immunoassay(s):
SDS-PAGE ([ DESCRIPTION]
Effective Size Range: 10kDa to 70kDa.
Protein bands: 10 kDa, 14 kDa, 18 kDa, 22 kDa, 26 kDa, 33 kDa, 44 kDa and 70 kDa.
Double intensity bands: The 26 kDa, 18 kDa, 10 kDa bands are at double intensity to make location and size approximation of proteins of interest quick and easy.
Ready-to-use: No need to heat, dilute or add reducing agents before use.
[ STORAGE BUFFER]
62.5mM Tris-H3P04 (pH 7.5 at 25°C), 1 mM EDTA, 2% (w/v) SDS, 100mM DTT, 1 mM NaN3, 0.01 % (w/v) bromo-phenol blue and 33% (v/v) glycerol.
Accurate protein sizing on SDS-polyacrylamide gels and Western blots.
1. Allow the Marker to reach room temperature and thoroughly mix before use. This will ensure that any solids that may have precipitated at -20°C have returned to solution. Do not boil!
2. Load the following volumes of the Marker on a SDS-polyacrylamide gel.
[ LOADING VOLUMES]
3~5uL per well for mini gel; 7uL per well for large gel.
Use the same volumes for Western blotting applications. The loading volumes listed above are recommended for gels with a thickness of 0.75~1.0mm. The loading volume should be double for 1.5mm thick gels.
[ QUALITY CONTROL]
The protein marker, with molecular weight shifts of < or = 5% and minimal band broadening, are confirmed by migration in SDS-PAGE system. Electrophoresis of the Marker on a 13~18% Tris-glycine SDS-polyacrylamide gel resolves 8 individual bands.
1. The protein marker is conveniently packaged and ready to use. There is no need to heat, dilute or add reducing agents. Do not boil, which may cause band degradation.
2. The molecular weights of the proteins have a lot-to-Iot variation of ~5%.
3. These protein markers are for SDS-PAGE and should not to be used for native electrophoresis or a native gel. These markers are denatured and have SDS in the storage buffer.
4. Don\'t load too much protein. See recommended load volumes in the manual.
5. Check % gel that is being used if the lands miss. Depending on gel type and/or percentage, you may not see all of the bands. For example, one would not see the smallest bands of the standard on a very low % gel. A high % gel may not resolve the higher MW bands. Homogeneous low percentage gels are recommended for analysis of large proteins and high percentage gels for analysis of small proteins. In high percentage gels (13~18%) large proteins (22~70kDa) can separate, while in low percentage gels (4~8%) small proteins (14 and 10kDa) will migrate with the tracking dye. Longer transfer times or higher transfer voltages may be required for Western blotting of large (>100kDa) proteins.
6. If additional bands are observed in the gel image of the protein ladder, this might be caused by Dn oxidation in the storage buffer.
7. If proteins show a poor transfer, increase voltage, current or length of time for transfer, and pay attention to the SDS from the transfer buffer. SDS will cause the proteins to bind less efficiently to membranes because it disrupts the hydrophobic interaction between the membrane and the protein. If SDS is present in transfer buffer, make sure that there is no more than 0.02% in buffer.
8. Alcohol enhances hydrophobic binding to membrane. Not enough alcohol may prevent binding.
[ SHELF LIFE]
Storage: -20 degree C, 1 year at -20 degree C. If the product has been stored as directed for one year, its shelf life maybe extended by adding dithiothreitol (Dn) to approximately 50mM.)
Our knowledge of the role of proteins in cellular processes is continually evolving. Most proteins, including Erythropoietin (EPO) are typically involved in one or more signaling pathways or biological processes. Professionally manufactured recombinant proteins are increasingly becoming essential and commonplace tools for elucidating new knowledge about the role of proteins in both health and disease.