Darbepoetin

Darbepoetin

Erythropoietin

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  • Name :

    Darbepoetin

    Description:

    Darbepoetin-Alpha Human Recombinant

    Erythropoietin-Mutant, EPO-mutant, NESP, Novel Erythropoiesis Stimulating Protein, EPO Mutant.

    Product # :

    CYT-1263

    Price :

    Quantity :

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    Room Temp Icon

    Shipped at Room temp

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  • description
  • source
  • formulation
  • purity
  • biological activity
  • More Info

Description

Darbepoetin-alpha Human Recombinant is produced in Chinese hamster ovary (CHO) cells by recombinant DNA technology is a glycosylated polypeptide chain containing 165 amino acids and having a predicted molecular mass of 18,200 Dalton and apparent glycosylated molecular mass of 37-40kDa. Darbepoetin is purified by proprietary chromatographic techniques.

Source

Chinese Hamster Ovary Cells(CHO).

Formulation

Each mg of lyophilized Darbepoetin powder contains 20mM Phosphate Buffer, 0.15M NaCl pH-6.2 & 0.005% Tween-80.

Purity

Greater than 97.0% as determined by SDS-PAGE.

Biological Activity

The Specific Activity was measured by Normocyth -aemic mice and was found to be 1,000,000 IU/mg.

More Info

  • Introduction

    Darbepoetin is a mutant recombinant human EPO-Alpha protein re-engineered, containing 5 amino acid changes at N30, T32, V87, N88, T90. Darbepoetin exhibits longer half-life stimulating red blood cell production than Erythropoietin. Darbepoetin maintains hemoglobin more effectively compared to EPO-Alpha.

  • Synonyms

    NESP, Novel Erythropoiesis Stimulating Protein, EPO Mutant.

  • Physical Appearance

    Sterile Filtered White lyophilized (freeze-dried) powder.

  • Stability

    Lyophilized Darbepoetin although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution Darbepoetin should be stored at 4°C between 2-7 days and for future use below -18°C.For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).Please prevent freeze-thaw cycles.

  • Solubility

    It is recommended to reconstitute the lyophilized Darbepoetin in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.

  • Amino Acid Sequence

    APPRLICDSR VLERYLLEAK EAENITTGCN ETCSLNENIT VPDTKVNFYA WKRMEVGQQA VEVWQGLALL SEAVLRGQAL LVNSSQVNET LQLHVDKAVS GLRSLTTLLR ALGAQKEAIS PPDAASAAPL RTITADTFRK LFRVYSNFLR GKLKLYTGEA CRTGD.

  • Background

    What is the molecular weight/Mw of DARBEPOETIN Protein?
    DARBEPOETIN Protein has a total Mw of 38.5kDa.

    What is the source or expression system of DARBEPOETIN Protein?
    Chinese Hamster Ovary Cells(CHO).

    What is the Purity of DARBEPOETIN Protein?
    DARBEPOETIN Protein is >97% pure as determined by SDS-PAGE.

    What is the Biological Activity of DARBEPOETIN Protein?
    The Specific Activity was measured by Normocyth -aemic mice and was found to be 1,000,000 IU/mg.

    What is the amino acid sequence of DARBEPOETIN Protein?
    APPRLICDSR VLERYLLEAK EAENITTGCN ETCSLNENIT VPDTKVNFYA WKRMEVGQQA VEVWQGLALL SEAVLRGQAL LVNSSQVNET LQLHVDKAVS GLRSLTTLLR ALGAQKEAIS PPDAASAAPL RTITADTFRK LFRVYSNFLR GKLKLYTGEA CRTGD

    What applications can DARBEPOETIN Protein be used in?
    DARBEPOETIN Protein can probably be used in western blot, ELISA and Lateral Flow.

    What is the endotoxin level for DARBEPOETIN Protein?
    The endotoxin level is minimal, DARBEPOETIN Protein was purified using conventional chromatography techniques

ProSpec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

Darbepoetin

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Frequently asked questions

1. Which company offers Recombinant Growth Factors at competitive prices?

ProSpec offers vast array of Growth Factors such as EGF| IGF1 | Leptin| VEGF and much more at affordable descent prices to meet academic budgets
ProSpec provides High Purity Growth Factors for 30 years, purity exhibits 95% and more as visualized by SDS-PAGE gel.
Growth Factors are in stock for immediate delivery via FedEx which enables scientists and researchers to perform immediate experiments.
Growth Factors are shipped globally using ProSpec’s custom tailored Styrofoam insulated box and premium ice packs that ensure maximum protein stability during transit to your laboratory.

2. Who can produce custom Growth Factor for me?

ProSpec offers custom production of recombinant Growth Factors, managing the entire process from gene design and cloning to protein expression and fermentation. Harvest and cell breakage. Purification using standard chromatography, quality control testing, and formulation. Freeze-drying (lyophilization) is available as an option. For more information contact us at info@prospecbio.com

3. How long does it take to produce a custom Growth Factor

The production time line for a custom recombinant growth factor can take about 5-7 weeks.

4. Where can i find biologically active Growth Factors?

ProSpec offers biologically functional Growth Factors such as Prolactin | Erythropoietin | PDGF-BB which are rigorously tested in-vitro to provide researchers world wide BioActive Growth Factors.

5. What is the activity validation method for Growth Factors?

ProSpec uses cell-based bioassays that measures the biological activity, for example:
Activin-A is validated by Activin-responsive luciferase assay, SMAD2/3 phosphorylation.
CSF, G-CSF, GM-CSF, M-CSF are validated by Cytokine-dependent cell proliferation assay.
CTGF is validated by Fibroblast proliferation or migration assay.
EGF is validated by NIH3T3 or Balb/3T3 proliferation assay.
Epigen (EPGN) is validated by EGFR activation assay, cell proliferation.
Erythropoietin (EPO) is validated by TF-1 or UT-7 erythroid cell proliferation assay.
Fibroblast Growth Factors is validated by NIH3T3 proliferation assay.
IGF-1 and IGF-2 are validated by Cell proliferation assay (MCF-7, Saos-2, BALB/3T3).

6. Is the Growth Factor full length or partial?

The Growth Factor proteins are both full length and partial while majority of them are active mature proteins rather than a full-length precursor. ProSpec active mature proteins exhibits higher expression yields, better stability, Easier purification and equivalent biological activity as full length precursor.

7. Is the Growth Factor monomeric or dimeric form?

This depends on the Growth Factor. ProSpec Growth Factors are supplied in the native biologically active oligomeric state, which can be monomeric, dimeric, trimeric, or even multimeric since because receptor binding and biological activity often depend on correct oligomerization.
The EGF, Epigen, Fibroblast Growth Factors (FGF1, FGF2, FGF7/KGF)
HDGF, IGFBPs (IGFBP-1 to IGFBP-7), IGF2 are usually provided as monomer proteins though Activin-A and myostatin are supplied as homodimers.

8. What is the Growth Factor purity and how was it determined?

The Growth Factors purity ranges from high purity of ≥95% to premium grade ≥97%.
ProSpec uses SDS-PAGE Coomassie Blue Staining as the primary purity method which Visualizes protein bands. Some SDS-PAGE gels are stained with Silver Stain. Other analytical methods include RP-HPLC [ High-resolution purity determination ] , SEC-HPLC for the detections of aggregates and fragments and Western Blot to confirm Growth Factors identity.

9. Is the Growth Factor sequence verified?

The amino acid sequence verification is performed at DNA & protein level.
Sanger DNA Sequencing verifies the expression plasmid having correct coding.
LC-MS is used to verify the molecular weight of the final purified Growth Factor
Prospec also uses Edman N-terminal Sequencing to determine the amino-terminal sequence.

10. What is the endotoxin level in Growth Factors and how do you test LPS ?

ProSpec Growth Factors are tested for endotoxin also named LPS [ lipopolysaccharide ] which is found in the outer membrane of Gram-negative bacteria which can affect cytokine secretion, cell proliferation and stem cell.
ProSpec measures endotoxin using the Limulus Amebocyte Lysate [ LAL ] assay such as Gel-Clot.
ProSpec standard Endotoxin level for cytokines is usually <1 EU/ug.
What Does "EU" Mean? EU = Endotoxin Units

11. Is there host cell protein contamination?

There might be a minor existence of host-cell proteins post chromatography purification and quality-control procedures that greatly minimize these impurities. ProSpec tests host-cell protein levels by SDS-PAGE silver staining, or mass spectrometry.

12. Is the protein tested in vitro, in-vivo cell culture, stem cells, T cells, ELISA, western blot?

PoSpec Growth Factors proteins are usually tested for biological activity and identity, while some proteins are not though supported by literature and customer experience. Interleukins are tested in T-cell, NK-cell, PBMC, stem cell culture. Interferons are tested in Antiviral assays, immune-cell activation, TNFA proteins are tested in Apoptosis, inflammation and immune studies. ProSpec does not conduct in-vivo or animal injections experiments in house or our source.