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Human Erythropoietin/EPO Antibody

Catalog #: MAB2871
20 Citations 1 Review Datasheet / COA / SDS
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Detection of Erythropoietin/EPO by Western Blot
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Product Details
Citations (20)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Erythropoietin/EPO Antibody Summary

Species Reactivity
Human
Specificity
Recognizes an epitope within the first 26 amino acids at the NH2 terminus of human urinary and human recombinant EPO. Binds human and rat EPO but does not bind sheep, baboon, or dog EPO (1, 2).
Source
Monoclonal Mouse IgG2A Clone # AE7A5
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Erythropoietin/EPO synthetic peptide analog
Ala28-Thr53
Accession # P01588
Formulation
Supplied as a 0.2 μm filtered solution in PBS. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
Recombinant Human Erythropoietin/EPO (Ultrapure) (Catalog # 286-EP)
Immunoaffinity Purification
This antibody can be used with optimal binding of human EPO at pH 7-9 (1-3).
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Erythropoietin/EPO by Western Blot View Larger

Detection of Erythropoietin/EPO by Western Blot Analysis of the expression of EPO-IgG1 variants. Total soluble protein (TSP) extracts from N. benthamiana delta XTFT leaves expressing different EPO-Fc variants were analyzed by immunoblotting at 5 days post-infiltration (dpi) using (A) anti-EPO and (B) anti-hIgG antibodies. (C) EPO-Fc variants purified out of TSP with protein A were stained with Coomassie Brilliant Blue under reducing conditions (CBB). (D) Oligomerization of EPO-Fc variants was evaluated in 8% native PAGE of TSP non-reducing samples. Ponceau staining shows similar amounts of protein loaded. Protein size markers are shown in kilo Dalton (kDa). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34305971), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Erythropoietin/EPO by Western Blot View Larger

Detection of Erythropoietin/EPO by Western Blot Western blots of mock-treated and alpha -L-arabinofuranosidase-digested samples from rhEPO-producing Physcomitrella lines. Ten microgram total protein of precipitated culture supernatants of the rhEPO-producing lines 174.16, delta galt1 and X24 were digested with one unit of alpha -L-arabinofuranosidase ( alpha -Arafase), while control samples were treated equivalently but without alpha -L-arabinofuranosidase (mock). After separation on SDS-PAGE and blotting, the PVDF-membrane was subsequently incubated with the anti-1,5-alpha -L-arabinan antibody (LM6-M, 1:10) (A) and an anti-hEPO monoclonal antibody (1:4,000) (B). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35252146), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Erythropoietin/EPO by Western Blot View Larger

Detection of Erythropoietin/EPO by Western Blot Western blots of precipitated culture supernatants of rhEPO-producing Physcomitrella lines. Five microgram total protein of the precipitated and blotted culture supernatants of the rhEPO-producing lines 174.16, delta galt1 and X24 were subsequently immunodetected with the anti-1,5-alpha -L-arabinan antibody (LM6-M, 1:10) (A) and the anti-hEPO monoclonal antibody (1:4,000) (B). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35252146), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Erythropoietin/EPO by Western Blot View Larger

Detection of Erythropoietin/EPO by Western Blot STABILON significantly boosts the intracellular and secreted levels of hEPO in mammalian cells. (a) Stably transfected CHO-K1 cells expressing hEPO (lane 2), hEPO-Stab (lane 3) or hEPO-2 × Stab (lane 4) proteins were induced by doxycycline treatment for 24 h. Lane 1 is the control nontransfected cell. Total cell extracts were fractionated on SDS-PAGE. Expression of hEPO derivatives (unmodified and glycosylated forms, Form I–III) was analyzed by immunoblotting using anti-hEPO monoclonal antibody. Coomassie-stained gel shows the protein loading (left panel). (b) 10 µL of cell culture medium of doxycycline-induced control (lane 1), hEPO (lane 2) or hEPO-Stab (lane 3) expressing stably transfected CHO-K1 cells were fractionated on SDS-PAGE. Secreted hEPO glycosylated species (Form III) were analyzed by immunoblotting. Coomassie-stained gel serves as loading control (left panel). Asterisk labels the extremely abundant serum albumin present in the medium. (c) Stable transfected CHO-K1 cells expressing hEPO, hEPO-Stab or hEPO-2 × Stab were induced with doxycycline treatment for 5 h, then continued for an additional 5 h in the presence of 0 μM (lanes 2, 5 and 8), 5 μM (lanes 3, 6 and 9) and 10 μM (lanes 4, 7 and 10) of MG132. Accumulation of hEPO derivatives was analyzed in total cell extracts by immunoblotting. Protein extracts of CHO-K1 cells served as control (lane 1). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35897744), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Erythropoietin/EPO by Western Blot View Larger

Detection of Erythropoietin/EPO by Western Blot STABILON significantly boosts the intracellular and secreted levels of hEPO in mammalian cells. (a) Stably transfected CHO-K1 cells expressing hEPO (lane 2), hEPO-Stab (lane 3) or hEPO-2 × Stab (lane 4) proteins were induced by doxycycline treatment for 24 h. Lane 1 is the control nontransfected cell. Total cell extracts were fractionated on SDS-PAGE. Expression of hEPO derivatives (unmodified and glycosylated forms, Form I–III) was analyzed by immunoblotting using anti-hEPO monoclonal antibody. Coomassie-stained gel shows the protein loading (left panel). (b) 10 µL of cell culture medium of doxycycline-induced control (lane 1), hEPO (lane 2) or hEPO-Stab (lane 3) expressing stably transfected CHO-K1 cells were fractionated on SDS-PAGE. Secreted hEPO glycosylated species (Form III) were analyzed by immunoblotting. Coomassie-stained gel serves as loading control (left panel). Asterisk labels the extremely abundant serum albumin present in the medium. (c) Stable transfected CHO-K1 cells expressing hEPO, hEPO-Stab or hEPO-2 × Stab were induced with doxycycline treatment for 5 h, then continued for an additional 5 h in the presence of 0 μM (lanes 2, 5 and 8), 5 μM (lanes 3, 6 and 9) and 10 μM (lanes 4, 7 and 10) of MG132. Accumulation of hEPO derivatives was analyzed in total cell extracts by immunoblotting. Protein extracts of CHO-K1 cells served as control (lane 1). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35897744), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Erythropoietin/EPO by Western Blot View Larger

Detection of Erythropoietin/EPO by Western Blot The effect of STABILON derivatives on the accumulation of hEPO in mammalian cells. (a) hEPO (lane 2), hEPO-StabDm1−15 (lane 3), hEPO-StabDm1−13 (lane 4), hEPO-StabDm1−10 (lane 5), hEPO-StabDm1−9 (lane 6), hEPO-StabDm1−8 (lane 7), hEPO-StabDm8−10 (lane 8), hEPO-StabDm8−11 (lane 9), hEPO-StabDm8−12 (lane 10), hEPO-StabDm1−7 (lane 11), hEPO-StabDm1−5 (lane 12), hEPO-StabDm1−3 (lane 13), hEPO-StabDm1−13-KR (lane 14), hEPO-StabDm1−13-KA (lane 15) and hEPO-StabDm1−13-DA (lane 16) transgenes in stably transfected CHO-K1 cells were induced by doxycycline-treatment for 24 h, respectively. The intracellular accumulation of hEPO derivatives was analyzed in total cell extracts by Western blotting using anti-hEPO monoclonal antibody. alpha Tubulin served as loading control. The lower diagram shows the amino acid positions and composition of the STABILON derivatives fused to hEPO. (b) hEPO (lane 2), hEPO-StabDm1-13 (lane 3) or hEPO-StabHs1–13 (lane 4) were expressed in stably transfected CHO-K1 cells by doxycycline-treatment for 24 h. Accumulation of hEPO derivatives was analyzed in total cell extracts by Western blotting using anti-hEPO monoclonal antibody. Protein extracts of CHO-K1 cells serve as control (lane 1). (c) Stably transfected CHO-K1 cells carrying the hEPO (lane 2), hEPO-StabDm1−13 (lane 3) and hEPO-StabHs1−13 (lane 4) genes were induced by doxycycline for 24 h. hEPO derivatives secreted into the tissue culture medium were analyzed by Western blotting with anti-hEPO monoclonal antibody. Tissue culture medium of CHO-K1 cells served as control (lane 1). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35897744), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Erythropoietin/EPO by Western Blot View Larger

Detection of Erythropoietin/EPO by Western Blot The effect of STABILON derivatives on the accumulation of hEPO in mammalian cells. (a) hEPO (lane 2), hEPO-StabDm1−15 (lane 3), hEPO-StabDm1−13 (lane 4), hEPO-StabDm1−10 (lane 5), hEPO-StabDm1−9 (lane 6), hEPO-StabDm1−8 (lane 7), hEPO-StabDm8−10 (lane 8), hEPO-StabDm8−11 (lane 9), hEPO-StabDm8−12 (lane 10), hEPO-StabDm1−7 (lane 11), hEPO-StabDm1−5 (lane 12), hEPO-StabDm1−3 (lane 13), hEPO-StabDm1−13-KR (lane 14), hEPO-StabDm1−13-KA (lane 15) and hEPO-StabDm1−13-DA (lane 16) transgenes in stably transfected CHO-K1 cells were induced by doxycycline-treatment for 24 h, respectively. The intracellular accumulation of hEPO derivatives was analyzed in total cell extracts by Western blotting using anti-hEPO monoclonal antibody. alpha Tubulin served as loading control. The lower diagram shows the amino acid positions and composition of the STABILON derivatives fused to hEPO. (b) hEPO (lane 2), hEPO-StabDm1-13 (lane 3) or hEPO-StabHs1–13 (lane 4) were expressed in stably transfected CHO-K1 cells by doxycycline-treatment for 24 h. Accumulation of hEPO derivatives was analyzed in total cell extracts by Western blotting using anti-hEPO monoclonal antibody. Protein extracts of CHO-K1 cells serve as control (lane 1). (c) Stably transfected CHO-K1 cells carrying the hEPO (lane 2), hEPO-StabDm1−13 (lane 3) and hEPO-StabHs1−13 (lane 4) genes were induced by doxycycline for 24 h. hEPO derivatives secreted into the tissue culture medium were analyzed by Western blotting with anti-hEPO monoclonal antibody. Tissue culture medium of CHO-K1 cells served as control (lane 1). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35897744), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

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Preparation and Storage

Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C, as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
  • 6 months, -20 to -70 °C under sterile conditions after opening.

Background: Erythropoietin/EPO

Erythropoietin (EPO) a glycoprotein produced primarily by the kidney, is the principal factor that regulates erythropoiesis by stimulating the proliferation and differentiation of erythroid progenitor cells.

Entrez Gene IDs
2056 (Human); 13856 (Mouse); 24335 (Rat)
Alternate Names
ECYT5; EP; EPO; epoetin; Erythropoietin; MGC138142; MVCD2

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Citations for Human Erythropoietin/EPO Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

20 Citations: Showing 1 - 10
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  1. Identification and single-base gene-editing functional validation of a cis-EPO variant as a genetic predictor for EPO-increasing therapies
    Authors: Charli E. Harlow, Josan Gandawijaya, Rosemary A. Bamford, Emily-Rose Martin, Andrew R. Wood, Peter J. van der Most et al.
    The American Journal of Human Genetics
  2. STABILON, a Novel Sequence Motif That Enhances the Expression and Accumulation of Intracellular and Secreted Proteins
    Authors: Z Rethi-Nagy, E Abraham, K Udvardy, E Klement, Z Darula, M Pal, RL Katona, V Tubak, T Pali, Z Kota, R Sinka, A Udvardy, Z Lipinszki
    International Journal of Molecular Sciences, 2022-07-25;23(15):.
    Species: Chinese Hamster, Drosophila
    Sample Types: Cell Lysates, Tisssue Homogenates
    Applications: Western Blot
  3. Unexpected Arabinosylation after Humanization of Plant Protein N-Glycosylation
    Authors: Lennard L. Bohlender, Juliana Parsons, Sebastian N. W. Hoernstein, Nina Bangert, Fernando Rodríguez-Jahnke, Ralf Reski et al.
    Frontiers in Bioengineering and Biotechnology
  4. The Instability of Dimeric Fc-Fusions Expressed in Plants Can Be Solved by Monomeric Fc Technology
    Authors: Pia Gattinger, Shiva Izadi, Clemens Grünwald-Gruber, Somanath Kallolimath, Alexandra Castilho
    Frontiers in Plant Science
  5. Modified recombinant human erythropoietin with potentially reduced immunogenicity
    Authors: T Susantad, M Fuangthong, K Tharakaram, P Tit-Oon, M Ruchirawat, R Sasisekhar
    Scientific Reports, 2021-01-15;11(1):1491.
    Species: Human
    Sample Types: Protein
    Applications: ELISA Capture
  6. In vivo modification of the goat mammary gland glycosylation pathway
    Authors: MJ Leiva-Carr, S Jiménez-Ch, DJ Harvey, NC Parra, KC Tavares, F Camacho, A González, O Sánchez, R Montesino, JR Toledo
    N Biotechnol, 2020-11-04;61(0):11-21.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  7. Erythropoietin production by the kidney and the liver in response to severe hypoxia evaluated by Western blotting with deglycosylation
    Authors: Y Yasuoka, T Fukuyama, Y Izumi, Y Nakayama, H Inoue, K Yanagita, T Oshima, T Yamazaki, T Uematsu, N Kobayashi, Y Shimada, Y Nagaba, M Mukoyama, T Yamashita, Y Sato, JM Sands, K Kawahara, H Nonoguchi
    Physiol Rep, 2020-06-01;8(12):e14485.
    Species: Human
    Sample Types: Plasma
    Applications: Western Blot
  8. Physicochemical and biological evaluation of JR-131 as a biosimilar to a long-acting erythropoiesis-stimulating agent darbepoetin alfa
    Authors: J Tani, Y Ito, S Tatemichi, M Yamakami, T Fukui, Y Hatano, S Kakimoto, A Kotani, A Sugimura, K Mihara, R Yamamoto, N Tanaka, K Minami, K Takahashi, T Hirato
    PLoS ONE, 2020-04-17;15(4):e0231830.
    Species: Human
    Sample Types: Protein
    Applications: Western Blot
  9. A human expression system based on HEK293 for the stable production of recombinant erythropoietin
    Authors: CL Chin, JB Goh, H Srinivasan, KI Liu, A Gowher, R Shanmugam, HL Lim, M Choo, WQ Tang, AH Tan, T Nguyen-Khu, MH Tan, SK Ng
    Sci Rep, 2019-11-14;9(1):16768.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  10. Generation of transgenic chickens expressing the human erythropoietin (hEPO) gene in an oviduct-specific manner: Production of transgenic chicken eggs containing human erythropoietin in egg whites
    Authors: MS Kwon, BC Koo, D Kim, YH Nam, XS Cui, NH Kim, T Kim
    PLoS ONE, 2018-05-30;13(5):e0194721.
    Species: Chicken
    Sample Types: Protein
    Applications: Western Blot
  11. Far-western blotting as a solution to the non-specificity of the anti-erythropoietin receptor antibody
    Authors: Barbora Fecková, Patrícia Kimáková, Lenka Ilkovičová, Erika Szentpéteriová, Nataša Debeljak, Zuzana Solárová et al.
    Oncology Letters
  12. Safety and angiogenic effects of systemic gene delivery of a modified erythropoietin.
    Authors: de Lucas Cerrillo A, Bond W, Rex T
    Gene Ther, 2015-02-26;22(5):365-73.
    Species: Hamster
    Sample Types: Whole Cells
    Applications: Functional Assay
  13. EPO gene expression promotes proliferation, migration and invasion via the p38MAPK/AP-1/MMP-9 pathway by p21WAF1 expression in vascular smooth muscle cells.
    Authors: Park S, Won S, Song J, Kambe T, Nagao M, Kim W, Moon S
    Cell Signal, 2014-12-08;27(3):470-8.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  14. Novel human renal proximal tubular cell line for the production of complex proteins.
    Authors: Fliedl L, Manhart G, Kast F, Katinger H, Kunert R, Grillari J, Wieser M, Grillari-Voglauer R
    J Biotechnol, 2014-02-16;176(0):29-39.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  15. The nexus between VEGF and NFkappaB orchestrates a hypoxia-independent neovasculogenesis.
    Authors: DeNiro M, Al-Mohanna F, Alsmadi O, Al-Mohanna F
    PLoS ONE, 2013-03-22;8(3):e59021.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  16. Generation of Biologically Active Multi-Sialylated Recombinant Human EPOFc in Plants
    Authors: Alexandra Castilho, Laura Neumann, Pia Gattinger, Richard Strasser, Karola Vorauer-Uhl, Thomas Sterovsky et al.
    PLoS ONE
    Species: Plant - Nicotiana benthamiana
    Sample Types: Protein
    Applications: Western Blot
  17. Engineering of Sialylated Mucin-type O-Glycosylation in Plants*
    Authors: Alexandra Castilho, Laura Neumann, Sasha Daskalova, Hugh S. Mason, Herta Steinkellner, Friedrich Altmann et al.
    Journal of Biological Chemistry
  18. Location and the functionality of erythropoietin receptor(s) in A2780 cells
    Authors: Peter Solár, Gabriela Hrčková, Lenka Varinská, Zuzana Solárová, Ján Kriška, Ivana Uhrínová et al.
    Oncology Reports
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  19. Quantifying Western blots: pitfalls of densitometry.
    Authors: Gassmann M, Grenacher B, Rohde B, Vogel J
    Electrophoresis, 2009-06-01;30(11):1845-55.
    Species: Bovine, Camelid, Canine, Equine, Feline, Human, Mouse, Porcine, Primate - Pongo (Orangutan), Proboscine (Elephant), Rat, Rhinoceros
    Sample Types: Plasma
    Applications: Western Blot
  20. Testing for recombinant erythropoietin.
    Authors: Delanghe JR, Bollen M, Beullens M
    Am. J. Hematol., 2008-03-01;83(3):237-41.
    Species: Human
    Sample Types: Urine
    Applications: Western Blot

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Human Erythropoietin Antibody
By Leslie Priddy on 04/13/2018
Application: IHC Sample Tested: Serum Species: Human