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In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
Cell lines
2 wild-type (wt) p53 (Lovo and LS174T) and 4 mutant (mt) p53 (WiDr, WiDr/F, HT29 and SW948) colon carcinoma cell lines
Preparation method
The solubility of this compound in DMSO is > 154mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below - 20 °C for several months.
Reacting condition
50 and 100 nM; 24 and 48 hrs
Applications
Raltitrexed up-regulated p53 and p21 expression in wt p53 cells, but not in mt p53 cells. In the mt p53 cells HT29 and WiDr/F, the highest induction of thymidylate synthase (6 ~ 10 folds) was observed after Raltitrexed treatment. Moreover, Raltitrexed increased Bax expression up to 5 folds in wt p53 cells, but with only a very slight induction of Bax expression in mt p53 cells. In wt p53 cells, Raltitrexed treatment hardly changed Bcl-2 expression.
Animal models
C57BL/6J-ApcMin/+ mice
Dosage form
3 or 5 mg/kg; twice a week or five times a week
In C57BL/6J-ApcMin/+ mice, Raltitrexed (3 mg/kg; twice a week) increased the average tumor number in the small intestine by 4 folds. When the dose of Raltitrexed was elevated to 5 mg/kg, five times a week, it resulted in a 10-fold increase in the average tumor number. Under all administration schedules, Raltitrexed was well-tolerated with only few treatment-related deaths occurring. Raltitrexed-induced tumors commonly occurred in the duodenum and jejunum, with few in the ileum and none in the colon.
Other notes
Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.
References:
[1]. Peters GJ, van Triest B, Backus HH, Kuiper CM, van der Wilt CL, Pinedo HM. Molecular downstream events and induction of thymidylate synthase in mutant and wild-type p53 colon cancer cell lines after treatment with 5-fluorouracil and the thymidylate synthase inhibitor raltitrexed. Eur J Cancer. 2000 May;36(7):916-24.
[2]. Murphy JT, Tucker JM, Davis C, Berger FG. Raltitrexed increases tumorigenesis as a single agent yet exhibits anti-tumor synergy with 5-fluorouracil in ApcMin/+ mice. Cancer Biol Ther. 2004 Nov;3(11):1169-76.