PDF《ANTI-THROMBOPOIETIN (TPO)》

ANTI-THROMBOPOIETIN (TPO) Developed in Goat IgG Fraction of Antiserum Product Number T

4805 Product Description Anti-Human Thrombopoietin is developed in goats using recombinant human Tpo, expressed in the insect cell line Sf 21, as the immunogen.

The product is purified by Protein G affinity chromatography. Tpo, the ligand for the receptor encoded by the c-Mpl proto-oncogene, acts as a stimulator of the development of megakaryocyte precursors of platelets. Similar to erythropoietin, Tpo leads to an increase in the number of circulating platelets. Tpo affects the entire thrombopoietic process, with stronger effects in the later stages. Other thrombopoietic cytokines include stem cell factor (SCF), IL-3, IL-6, and IL-11. Tpo is an approximately

35 kDa polypeptide of

335 amino acids. However, due to glycosylation the protein has an apparent molecular weight of

75 kDa in SDS- PAGE. The precursor form of Tpo consists of

356 amino acids. To generate the mature Tpo (335 amino acids), the precursor cleaves a

21 amino acid signal peptide. Human, mouse and dog Tpo shows 69-75% amino acid homology. Reagents Goat Anti-Human Tpo is provided lyophilized from phosphate buffered saline, to which no preservatives are added. Reconstitution To one vial of lyophilized powder, add

1 ml of 0.2 ?m-filtered PBS to produce a

1 mg/ml stock solution of Anti-Human Tpo. If aseptic technique is used, no further filtration should be needed for use in cell culture environments. Storage/Stability Prior to reconstitution, store at C20 °C. Reconstituted product may be stored at 2-8 °C for up to one month. For prolonged storage, freeze in working aliquots at C20 °C. Avoid repeated freezing and thawing. Product Profile Anti-Human Thrombopoietin is tested for its ability to neutralize the biological activity of recombinant human Tpo on the human cell line, MO7e. The ND50 of the antibody is defined as the concentration of antibody resulting in a one-half maximal inhibition of bioactivity of recombinant human Tpo, when recombinant human Tpo is present at a concentration just high enough to elicit a maximum response. In this bioassay,

10 ng/ml rhTpo was mixed with various dilutions of the antibody for

1 hour at

37 °C. After preincubation, the antigen- antibody mixture was added to MO7e cells in a 96-well plate. The assay mixture was incubated at

37 °C for

72 hours in a humidified CO2 incubator and pulsed for the final

4 hours with

3 H-thymidine. Cells were harvested onto glass filters and the

3 H-thymidine incorporation into DNA was measured. The antibody may also be used in immunoblotting and ELISA. In addition, the antibody does not cross-react with other cytokines tested.* * rhANG, rhAR, rhB7-1, rhB7-2, rmB7-2, rhBTC, rhβ-NGF, rhBDNF, rmC10, rhCD4, rhCD8, rhCD28, rhCNTF, rrCNTF, rhEGF, rhENA-78, rhEpo, rhFGFa, rhFGFb, rhFGF-4, rhFGF-5, rhFGF-6, rhFGF-7, rhFGF-9, rhG-CSF, rmG-CSF, rhGM-CSF, rhGM-CSF Rα, rmGM-CSF, rhGROα, rhGROβ, rhGROγ, rhHB-EGF, rhHRG-α, rhHGF, rhI-309, rhIFN-γ, rhIGF-I, rhIGF-I R, rhIGF-II, rhIL-1α, rhIL-1 RI, rhIL-1 RII, rmIL-1α, rhIL-1β, rmIL-1β, rhIL-1 ra, rmIL-1 ra, rhIL-2, rhIL-2 sRα, rhIL-2 sRβ, rhIL-2 sRγ, rmIL-2, rhIL-3, rhIL-3 sRα, rmIL-3, rhIL-4, rhIL-4 sR, rmIL-4, rhIL-5, rhIL-5 sRα, rhIL-5 sRβ, rmIL-5, rhIL-5, rhIL-6, rhIL-6 sR, rmIL-6, rhIL-7, rhIL-7 R, rmIL-7, rhIL-8, rhIL-9, rmIL-9, rhIL-10, rhIL-10 sR, rmIL-10, rhIL-11, rhIL-12, rmIL-12, rhIL-13, rmIL-13, rhIL-15, rhIP-10, rhJAK-1, rmJAK-1, rmJE, rhLIF, rhLIF R, rmLIF, rhM-CSF, rmM-CSF, rhMCP-1, rhMCP-1 R, rhMCP-2, rhMCP-3, rhMidkine, rhMIP-1α, rmMIP-1α, rhMIP-1β, rmMIP-1β, rmMIP-2, rhNT-3, rhNT-4, rhOSM, rhPD-ECGF, hPDGF, pPDGF, rhPDGF-AA, rhPDGF-AB, rhPDGF-BB, rhPDGF Rα, rhPlGF, rhPTN, rhRANTES, rhSCF, rmSCF, rhsgp130, rhSLPI, rhSTAT-1, rmSTAT-4, hTfR, rhTGF-α, rhTGF-β1, rhTGF-β2, rhTGF-β3, raTGF-β5, rhLAP (TGF-β1), rhLatent TGF-β1, rhTGF-β sRII, rhTGF-β sRIII, rhTNF-α, rmTNF-α, rhTNF-β, rhsTNF RI, rhsTNF RII, rhVEGF. JWM/KMR 06/02 Sigma brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip.