甲状旁腺激素(PTH)活性蛋白

[ PROPERTIES ]

Source: Prokaryotic expression. Host: E. coli

Residues: Ala32~Gln115

Tags: Two N-terminal Tags, His-tag and MBP-tag

Purity: >92%

Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.05% sarcosyl

and 5% trehalose. Applications: Cell culture; Activity Assays; In vivo assays. (May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 5.8

Predicted Molecular Mass: 59.4kDa

Accurate Molecular Mass: 59kDa as determined by SDS-PAGE reducing conditions.

[ USAGE ]

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/mL. Do not vortex. 

[ STORAGE AND STABILITY ]

Storage: Avoid repeated freeze/thaw cycles. 

Store at 2-8^oC for one month. Aliquot and store at -80^oC for 12 months.

Stability Test: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37^oC for 48h, and no obvious degradation and precipitation wereobserved. The loss rate is less than 5% within the expiration date underappropriate storage condition. 

[ SEQUENCE ]

[ ACTIVITY ] 

PTH (Parathyroid hormone) is a hormone secreted by the parathyroid glands thatis important in bone remodeling. As reported, osteoblast-like cell lines, such asROS 17/2.8, UMR106, SaOS, U2OS, MG63, that exhibit PTHR1, respond withincreased proliferation to PTH. Rat PTH shares similarities with human PTH inamino acid sequence with the identity of 71.3%. Thus, a proliferation assay of ratrecombinant PTH was conducted using U2OS cells. Briefly, U2OS cells wereseeded into triplicate wells of 96-well plates at a density of 2,000 cells/well andallowed to attach overnight, then the medium was replaced with serum-freestandard DMEM prior to the addition of various concentrations of PTH. Afterincubated for 48h, cells were observed by inverted microscope and cellproliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10µL of CCK-8solution was added to each well of the plate, then the absorbance at 450nm wasmeasured using a microplate reader after incubating the plate for 1-4 hours at37°C. Proliferation of U2OS cells after incubation with PTH for 48h observed byinverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8(Cell Counting Kit-8 ) assay after incubation with recombinant PTH for 48h. Theresult was shown in Figure 2. It was obvious that PTH increased cell viability ofU2OS cells.

[ IDENTIFICATION ]

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