Erythropoietin R Antibody (38409) [Unconjugated]

Western blot shows lysates of UT-7 human acute myeloid leukemia cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Erythropoietin R Monoclonal Antibody (Catalog # MAB307) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Erythropoietin R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-322-PB) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Erythropoietin R at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

TF-1 human erythroleukemic cell line was stained with Mouse Anti-Human Erythropoietin R Monoclonal Antibody (Catalog # MAB307, filled histogram) or isotype control antibody (MAB0041, open histogram) followed by anti-Mouse IgG Allophycocyanin-conjugated Secondary Antibody (F0101B). Staining was performed using our Staining Membrane-associated Proteins protocol.

sEpoR characterization in uremic serum.1a. Soluble EpoR is detectable in serum from dialysis patients by western blot. Human serum was subjected to immunoprecipitation with goat anti-human erythropoietin receptor antibody (R&D Systems, AF-322-PB) followed by western blotting with mouse monoclonal anti-human erythropoietin receptor (R&D Systems, MAB307). Both antibodies recognize the extracellular domain of the receptor. Lanes 1–6 are serum from 6 representative dialysis patients, lane 7 is blank and lane 8 is recombinant sEpoR (Sigma Aldrich E0643, Saint Louis MI). Shown in the serum samples is a band of expected molecular weight of approximately 27 kDa. The control sEpoR with Fc tag is consistent with the manufacturers reported molecular weight of 32 kDa. 1b. Soluble EpoR is also detected using the same dialysis patient serum samples by performing immunoprecipitation in reverse order. In this experiment immunoprecipitation was done with mouse monoclonal anti-human erythropoietin receptor (R&D Systems, MAB307) followed by western blotting with goat anti-human erythropoietin receptor (R&D Systems, AF-322-PB). Lanes 1 to 3 are serum from 3 dialysis patients, and lane 4 is recombinant sEpoR-Fc (Sigma, 307) as positive control. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20169072), licensed under a CC-BY license. Not internally tested by R&D Systems.

The 59 kDa protein detected by M-20 In MCF-7 cells is not bound by other anti-EpoR antibodies.The indicated lysates were immunoprecipitated (IP) then the immunoblotted (IB) with the indicated antibodies: ab10653 (abcam Inc), Mab307 (R&D systems), C-20 & M-20 (Santa Cruz Inc) or A-82 (Amgen Inc). COS cell lysates expressing a FLAG-tagged version of EpoR (FLAG-EpoR) [6] and UT-7/Epo cells served as EpoR positive controls. 769-P cells served as the EpoR negative control. (A) Westerns were immunoprecipitated (IP) with ab10653 or M-20 followed by immunoblotting (IB) with M-20. The position of full-length 59 kDa EpoR in positive controls is indicated by the arrow. Positions of molecular weight markers (kDa) are shown. Bands detected in 769-P lysates are non-EpoR cross-reacting proteins and include antibody chains that were not removed completely or protein G that leached from beads. Note the detection of a 59 kDa band with MCF-7 cells with the M:20/M:20 combination but not with the ab10653/IB:M-20 combination. (B) IP:IB combinations with the indicated antibodies were subjected to western analysis. The western slice containing the 59 kDa EpoR band from each combination is shown. Note the 59 kDa bands detected in EpoR positive controls but not 769-P cells. Only the M-20:M-20 combination detected a 59 kDa band in MCF-7 cells. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0068083), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Flow, IP
• Clone: 38409
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Erythropoietin R Antibody (38409) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Erythropoietin R
  Pro26-Pro250
  Accession # P19235
• Specificity: Detects human Erythropoietin R in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant mouse Erythropoietin R is observed.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: EPOR
• Purity Statement: Protein A or G purified from ascites

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunoprecipitation
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• 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 reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Erythropoietin R Antibody (38409) [Unconjugated]

• EpoR
• EPO-R
• Erythropoietin R
• erythropoietin receptor
• MGC138358

Background

Epo R is a transmembrane protein expressed on the surface of megakaryocytes, erythroid progenitors, and endothelial cells. It binds Epo and transmits signals that stimulate the proliferation and maturation of bone marrow erythroid precursors into red cells.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Erythropoietin R Antibody (MAB307)(8)

Publications using MAB307

• Patterson , Scott D, Rossi , John M, Paweletz , Katherin, Fitzpatrick , V Dan, Begley , C Glenn, Busse , Leigh, Elliott , Steve, McCaffery , Ian Functional EpoR pathway utilization is not detected in primary tumor cells isolated from human breast, non-small cell lung, colorectal, and ovarian tumor tissues. PLoS ONE, 2015-03-25;10(3):e0122149. 2015-03-25 [PMID: 25807104] (Flow Cytometry, Human)
• Anita Rózsás, Judit Berta, Lívia Rojkó, László Z. Horváth, Magdolna Keszthelyi, István Kenessey et al. Erythropoietin Receptor Expression Is a Potential Prognostic Factor in Human Lung Adenocarcinoma PLoS ONE 10/14/2013 [PMID: 24155958]
• Séverine Lamon, Aaron P. Russell The role and regulation of erythropoietin (EPO) and its receptor in skeletal muscle: how much do we really know? Frontiers in Physiology 7/15/2013 [PMID: 23874302]
• Steve Elliott, Susan Swift, Leigh Busse, Sheila Scully, Gwyneth Van, John Rossi et al. Epo Receptors Are Not Detectable in Primary Human Tumor Tissue Samples PLoS ONE 7/4/2013 [PMID: 23861852]
• De Grandis , Maria, Cambot , Marie, Wautier , Marie-Pa, Cassinat , Bruno, Chomienne , Christin, Colin , Yves, Wautier , Jean-Luc, Le Van Kim , Caroline, El Nemer , Wassim JAK2V617F activates Lu/BCAM-mediated red cell adhesion in polycythemia vera through an EpoR-independent Rap1/Akt pathway. Blood, 2012-11-16;121(4):658-65. 2012-11-16 [PMID: 23160466] (Flow Cytometry, Human)
• Thilo Welsch, Stefanie Zschäbitz, Verena Becker, Thomas Giese, Frank Bergmann, Ulf Hinz et al. Prognostic Significance of Erythropoietin in Pancreatic Adenocarcinoma PLoS ONE 8/1/2011 [PMID: 21829709]
• Khankin EV, Mutter WP, Tamez H Soluble erythropoietin receptor contributes to erythropoietin resistance in end-stage renal disease. PLoS ONE, 2010-02-16;5(2):e9246. 2010-02-16 [PMID: 20169072] (Immunoprecipitation, Western Blot, Human)
• Kirkeby A, van Beek J, Nielsen J, Leist M, Helboe L Functional and immunochemical characterisation of different antibodies against the erythropoietin receptor. J Neurosci Methods, 2007-04-12;164(1):50-8. 2007-04-12 [PMID: 17524492] (ICC, IHC, Western Blot, Human, Rat)

Bioinformatics

• Gene Symbol: EPOR
• Uniprot:
  • P19235()