Recombinant Human Erythropoietin Receptor

Recombinant Human Erythropoietin Receptor

Product No.: E225

[product_table name="All Top" skus="E225"]

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Alternate Names
EP, MGC138142
Product Type
Recombinant Protein
Expression Host
NS0 Cells
Species
Human

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Background

The erythropoietin receptor is a transmembrane protein belonging to the member of the cytokine receptor family. The receptor is tyrosine phosphorylated upon binding by erythropoietin and associates with and activates the tyrosine kinase, JAK2, which activates different intracellular pathways including: Ras/MAP kinase, phosphatidylinositol 3-kinase and STAT transcription factors. EPO-R transmits signals important for both proliferation and differentiation along the erythroid lineage.1 Defects in the erythropoietin receptor may produce erythroleukemia and familial erythrocytosis.

Protein Details

Purity
>90% by SDS-PAGE and analyzed by silver stain.
Endotoxin Level
<0.1 ng per 1 µg as determined by the LAL method
Biological Activity
The biological activity of Human Epo R was determined by its ability to inhibit Epodependent proliferation of TF1 human erythroleukemic cells. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323. The expected ED<sub>50</sub> is 1 - 4 ng/ml in the presence of 0.2 U/ml of rhEpo.
Fusion Protein Tag
Fc Fusion Protein
Protein Accession No.
Amino Acid Sequence
apppnlpdpk feskaallaa rgpeellcft erledlvcfw eeaasagvgp gnysfsyqle depwklcrlh qaptargavr fwcslptadt ssfvplelrv taasgapryh rvihinevvl ldapvglvar ladesghvvl rwlpppetpm tshiryevdv sagngagsvq rveilegrte cvlsnlrgrt rytfavrarm aepsfggfws awsepvsllt psdldpkscd kthtcppcpa pellggpsvf lfppkpkdtl misrtpevtc vvvdvshedp evkfnwyvdg vevhnaktkp reeqynstyr vvsvltvlhq dwlngkeykc kvsnkalpap iektiskakg qprepqvytl ppsrdeltkn qvsltclvkg fypsdiavew esngqpenny kttppvldsd gsfflysklt vdksrwqqgn vfscsvmhea lhnhytqksl slspgk
State of Matter
Lyophilized
Predicted Molecular Mass
The predicted molecular weight of Recombinant Human Epo R is Mr 51.4 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is Mr 55-60 kDa.
Predicted Molecular Mass
51.4
Formulation
This recombinant protein was 0.2 µm filtered and lyophilized from PBS with 0.1 mM EDTA and 0.05 % CHAPS, pH 7.2.
Storage and Stability
This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.
Country of Origin
USA
Shipping
Next Day Ambient
NCBI Gene Bank

Leinco Protein Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

Using Recombinant Human Erythropoietin Receptor (EPO-R) in research applications is essential for studying erythropoietin (EPO) signaling, receptor-ligand interactions, and downstream biological effects in hematopoiesis, tissue protection, and disease models.

Key scientific applications and advantages:

  • Mechanistic Studies: Recombinant EPO-R enables precise analysis of EPO binding, receptor activation, and subsequent intracellular signaling pathways (e.g., JAK2-mediated cascades), which are critical for understanding erythroid cell proliferation and differentiation.
  • Bioassays and ELISA Standards: Recombinant EPO-R is used as a standard in ELISA assays to quantify EPO or to validate receptor-ligand binding specificity and affinity in vitro.
  • Cellular and Tissue Models: EPO-R is expressed in various tissues (brain, retina, heart, kidney, endothelial cells, tumor cells), allowing investigation of EPO’s non-hematopoietic effects such as neuroprotection, anti-apoptotic activity, and tissue repair.
  • Drug Screening and Development: Recombinant EPO-R is valuable for screening EPO analogs, antagonists, or mimetics, facilitating the development of therapeutics targeting anemia, neurodegeneration, or ischemic injury.
  • Pathophysiological Research: Studying recombinant EPO-R helps elucidate its role in diseases such as cancer (e.g., EPO-R-mediated proliferation in hepatocellular carcinoma), ischemic injury, and chronic inflammation.

Best practices:

  • Use carrier-free recombinant EPO-R for biochemical assays or receptor-ligand binding studies to avoid interference from stabilizing agents.
  • For cell or tissue culture, select recombinant EPO-R preparations with carrier proteins (e.g., BSA) to enhance stability and reproducibility.

Summary of scientific rationale:

Employing recombinant EPO-R in research provides a controlled, reproducible system to dissect EPO signaling, model disease mechanisms, and evaluate therapeutic interventions across hematopoietic and non-hematopoietic contexts. This is critical for advancing understanding of erythropoiesis, tissue protection, and receptor pharmacology.

Yes, you can use recombinant human erythropoietin receptor (EpoR) as a standard for quantification or calibration in ELISA assays, provided the assay is designed to detect EpoR and the recombinant protein is compatible with your assay format.

Key considerations and supporting details:

  • Intended Use: Recombinant human EpoR is commonly used as a standard in ELISA assays that are specifically designed to quantify EpoR, not erythropoietin (EPO) itself. The product information for recombinant EpoR explicitly lists "ELISA (Standard)" as an application, indicating its suitability for this purpose.

  • Assay Type: Ensure your ELISA is for EpoR detection (not EPO). EpoR ELISAs use recombinant EpoR as a standard to generate a calibration curve for quantifying EpoR in samples. In contrast, EPO ELISAs use recombinant EPO as the standard.

  • Formulation: Recombinant EpoR is available in formulations with or without carrier proteins (e.g., BSA). For ELISA standards, the version with BSA is generally recommended for stability, unless BSA interferes with your assay.

  • Standard Preparation: Follow the manufacturer’s instructions for reconstitution and dilution to prepare the standard curve. Typically, the recombinant EpoR is reconstituted in PBS (with or without BSA) and serially diluted to generate the calibration curve.

  • Validation: Confirm that the recombinant EpoR standard is recognized equivalently to native EpoR by the antibodies used in your ELISA. Most commercial EpoR ELISA kits are validated to detect both natural and recombinant EpoR.

  • Documentation: Always refer to the technical datasheet for your specific recombinant EpoR and ELISA kit to ensure compatibility and optimal performance.

Summary Table: Use of Recombinant EpoR as ELISA Standard

ApplicationStandard UsedNotes
EpoR ELISARecombinant human EpoRSuitable as standard for quantification/calibration
EPO ELISARecombinant human EPONot interchangeable with EpoR standard

Important: Do not use recombinant EpoR as a standard in EPO quantification assays, as these require recombinant EPO, not its receptor.

If your ELISA is for EpoR detection, recombinant human EpoR is appropriate and widely used as a standard for quantification and calibration.

Recombinant Human Erythropoietin Applications in Published Research

Recombinant human erythropoietin (rHuEPO) has been validated for a diverse range of clinical and research applications across hematopoietic and non-hematopoietic domains.

Primary Hematopoietic Applications

The most established clinical applications involve treating various forms of anemia. rHuEPO is approved for anemia associated with chronic kidney disease, including both dialysis-dependent and non-dialysis patients. It is also validated for treating chemotherapy-induced anemia in patients with nonmyeloid malignancies, and for managing anemia in HIV-infected patients receiving zidovudine therapy. Additionally, rHuEPO has been validated for reducing allogeneic blood transfusion requirements in patients undergoing elective orthopedic surgery.

The mechanism underlying these applications involves rHuEPO's ability to promote erythropoiesis by enhancing survival of erythroid progenitor cells and primitive erythrocytes, thereby increasing red blood cell production and hemoglobin concentration.

Emerging Non-Hematopoietic Applications

Beyond traditional hematologic uses, rHuEPO has demonstrated therapeutic potential in multiple organ systems. Research has validated its application in neuroprotection and neuroregenerative therapy, with documented benefits in ischemic stroke and chronic schizophrenia shown in double-blind placebo-controlled clinical studies. Preclinical findings suggest therapeutic potential for neurological conditions including head trauma and epilepsy.

rHuEPO exhibits cytoprotective properties through anti-oxidative, anti-apoptotic, and anti-inflammatory mechanisms. These protective effects have led to investigation in cardiovascular disease, hepatobiliary surgery, and general surgery applications. The protein also shows promise in treating retinal diseases and other conditions affecting the nervous system.

Research and Bioactivity Validation

In laboratory settings, rHuEPO has been validated for bioactivity assessment and Western blot control applications, with bioactivity measured through its ability to inhibit erythropoietin-dependent cell proliferation in standardized assays.

Engineered Variants

Recent research has validated immunogenicity-reduced variants of rHuEPO designed through computational protein engineering, which maintain biological function while reducing immune responses in specific patient populations.

Reconstitution Protocol

Reconstitute the lyophilized recombinant human erythropoietin receptor (EPOR) protein by first centrifuging the vial briefly prior to opening to bring contents to the bottom. Reconstitute the protein in sterile distilled water to a concentration of 0.1–1.0 mg/mL. For optimal results, allow the vial and reconstitution buffer to equilibrate to room temperature before adding the buffer, then permit 15–30 minutes of gentle agitation for complete dissolution. If flakes persist, continue mixing for approximately 2 hours at room temperature.

Buffer and Formulation Considerations

The specific reconstitution buffer depends on your experimental requirements and the formulation type:

Standard formulations typically contain trehalose and carrier proteins (such as bovine serum albumin) and should be reconstituted in sterile PBS containing at least 0.1% human or bovine serum albumin.

Carrier-free formulations are reconstituted in sterile PBS without additional carrier proteins.

Alternative approach involves reconstituting in sterile distilled water alone, with optional addition of 5–50% glycerol (final concentration) to enhance stability.

Avoid vigorous vortexing or mixing of protein solutions, as this can denature the protein and reduce biological activity.

Storage and Stability

After reconstitution, aliquot the protein into working portions and store at ≤–20°C for long-term stability. Reconstituted protein solutions can be maintained at 4–8°C for 1–7 days, depending on formulation. Aliquots stored at –20°C remain stable for approximately 3 months.

Critical consideration: Avoid repeated freeze-thaw cycles, as these significantly compromise protein integrity and biological function. Use a manual defrost freezer and plan your aliquoting strategy accordingly to minimize thawing events.

Pre-Experimental Dilutions

For cell culture experiments requiring lower working concentrations, prepare further dilutions of the reconstituted stock in culture medium or buffered solutions containing carrier proteins, such as PBS with 0.1% bovine serum albumin. This approach protects the protein from non-specific adsorption to vessel surfaces and maintains biological activity during experimental procedures.

References & Citations

1. Siren, AL. et al. (2001) Proc Natl Acad Sci USA 98: 4044
2. Haroon, ZA. et al. (2003) Am J Pathol 163: 993

Certificate of Analysis

IMPORTANT Use lot specific datasheet for all technical information pertaining to this recombinant protein.
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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.