The thrombopoietin receptor also known as the myeloproliferative leukemia protein or CD110 (Cluster of Differentiation 110) is a protein that in humans is encoded by the MPL (myeloproliferative leukemia virus) oncogene. Thrombopoietin was shown to be the major regulator of megakaryocytopoiesis and platelet formation. The protein encoded by the c-mpl gene, CD110, is a 635 amino acid transmembrane domain, with two extracellular cytokine receptor domains and two intracellular cytokine receptor box motifs . TPO-R deficient mice were severely thrombocytopenic, emphasizing the important role of CD110 and thrombopoietin in megakaryocyte and platelet formation. Upon binding of thrombopoietin, CD110 is dimerized and the JAK family of non-receptor tyrosine kinases, as well as the STAT family, the MAPK family, the adaptor protein Shc and the receptors themselves become tyrosine phosphorylated.
Background References
1. Spivak JL et al. The Thrombopoietin Receptor, MPL, Is a Therapeutic Target of Opportunity in the MPN. Front Oncol. 2021 Mar
2. Vainchenker W et al. The role of the thrombopoietin receptor MPL in myeloproliferative neoplasms: recent findings and potential therapeutic applications. Expert Rev Hematol. 2019 Jun
Western blot analysis of TPOR / MPL on different lysates with Rabbit anti-TPOR / MPL antibody (HA500360) at 1/1,000 dilution.
Lane 1: A549 (Human lung adenocarcinoma cell) cell lysate Lane 2: Raji (Human Burkitt's lymphoma cell) cell lysate Lane 3: THP-1 (Human acute monoblastic leukemia cell) cell lysate
Lysates/proteins at 20 µg/Lane.
Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA500360, 1/1,000 in 5% NFDM/TBST, 2 hours at room temperature Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1:300,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 71 kDa Observed band size: 75 kDa
Immunocytochemistry analysis of THP-1 cells labeling TPOR / MPL with Rabbit anti-TPOR / MPL antibody (HA500360) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TPOR / MPL antibody (HA500360) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of SH-SY5Y cells labeling TPOR / MPL with Rabbit anti-TPOR / MPL antibody (HA500360) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-TPOR / MPL antibody (HA500360) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
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