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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.
Temozolomide, an alkylating agent, is a DNA damage inducer.
Cell lines
SK-LMS-1 leiomyosarcoma (MGMT-/p53+), Ewing sarcoma A-673 and GIST-T1 (both lines with MGMT+/p53- phenotype), and glioblastoma T98G (MGMT+/p53+)
Preparation method
The solubility of this compound in DMSO is >6.6 mg/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
62.5, 125, 250 and 500 μM; 72 h
Applications
In SK-LMS-1 cells, Temozolomide inhibited proliferative activity of SK-LMS-1 cells. A-673 cells was most sensitive to temozolomide, the effect was time- and dose-dependent. Preincubation of Ewing sarcoma cells with O6-benzylguanine potentiated the cytotoxic effect of the alkylating agent and reduced viability of tumor cells. GIST-T1 cells were insensitive to temozolomide.
Animal models
PARP1 wild‐type (WT) and PARP1 knock‐out (KO) mice
Dosage form
68 mg/kg; once daily for 5 days; orally administrated
Application
In PARP1 WT mice, temozolomide significantly lowered concentrations of NAD+ in the liver when compared with the control group (by 22%, p = 0.02). In the livers of PARP1 KO mice, there was also a statistically significant reduction in NAD+ in the temozolomide‐only group when compared with the control (by 22%, p = 0.03).
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] Khusnutdinov RR1, Boichuk SV2. Mechanisms of Sensitivity of Soft Tissue Sarcoma Cells to Temozolomide. Bull Exp Biol Med. 2017 Jul 18.
[2] Almeida GS1, Bawn CM1, Galler M1, et al. PARP inhibitor rucaparib induces changes in NAD levels in cells and liver tissues as assessed by MRS. NMR Biomed. 2017 May 22.
