2-Methylenebutyrolactone

Catalog No.S5148 Batch:S514801

Technical Data

Formula	C₅H₆O₂
Molecular Weight	98.10	CAS No.	547-65-9
Solubility (25°C)*	In vitro


* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Biological Activity

Description	2-Methylenebutyrolactone (Tulipalin A, MBL, α-methylene-γ-butyrolactone), also known as α-methylene-γ-butyrolactone (MBL) (Tulipalin A), belongs to the class of sesquiterpene lactone family and is considered as cyclic analog of most common vinyl monomer methyl methacrylate (MMA).
In vitro	After exposure of Jurkat T cells to 2-Methylenebutyrolactone (Tulipalin A , TUPA) for 72 h, a dose dependent toxicity is detected. Concentrations higher than 20 μM lead to a significant reduction of viable cells. Compared to vehicle-treated control cells, concentrations of 47.6 and 26.8 μM decrease the cell viability significantly by 50% (IC50) and 10% (IC10), respectively. In contrast, THP-1 cells are less sensitive toward TUPA. Concentrations >41 μM lead to a significant reduction of viable cells. IC10 is calculated with 50 μM, whereas a concentration of 83 μM is necessary to decrease the cell viability to 50% (IC50). In Jurkat T cells, four proteins responsible for the de novo purine synthesis, namely phosphoribosylformylglycinamidine synthase (PFAS), GMP synthase (GMPS), and ribosephosphate pyrophosphokinases 1 and 2 (PRPS1/2) are increased through TUPA treatment. Glutamine is also increased after TUPA treatment. In addition to the proteins belonging to the purine synthesis pathway, the abundances of proteins for DNA synthesis and repair (XRCC5, XRCC6, MCM3, MCM6, MCM7, TRA1) are also increased/induced during the treatment. While in THP-1 cells, no indication for higher purine synthesis induced by TUPA in subtoxic concentrations is discovered. TUPA induces the expression of proteins in Jurkat T cells responsible for cell stress, drug response, and protein folding: HYOU1, PDIA3, and DNAJB11. Additionally, the treatment with TUPA leads to the induction of three heat shock proteins (HSP90AA1, HSP90AB1, and HSP90B1) and three proteins belonging to the TRiC complex (CCT3,6,8) that are also responsible for proper protein folding. Treatment with TUPA leads to the induction of ROS that have adverse effects on Jurkat T cells and evokes slight cell stress responses in THP-1 cells. TUPA has influence on proteins in Jurkat T cells that contribute to different immune-specific, especially immunostimulating, reactions as allergic contact dermatitis. In THP-1 cells, no proteins regarding immune-specific reactions are up- or downregulated due to TUPA treatment^[1].
Density	1.119 g/mL at 25 °C

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines Human leukemic Jurkat T cells and human leukemic monocytes THP-1 cells Concentrations 10 to 82 μM (Jurkat cells); 20 to 163 μM (THP-1 cells) Incubation Time 72 h Method The influence of TUPA on the cell viability is measured using the MTT assay. Cells are seeded in flat bottom 96-well plates at a density of 4 × 10⁵ cells/mL. Afterward, Jurkat cells are exposed to TUPA in concentrations ranging from 10 to 82 μM. THP-1 cells (4 × 10⁵ cells/mL) are exposed to concentrations ranging from 20 to 163 μM. After incubation for 72 h under humidified conditions (5% CO2, 37℃), 10 μL of MTT (5 mg/mL) per well are added and cells are further incubated for 3 h. Subsequently, 200 μL DMSO are added for cell lysis and formazan solubilization following an incubation of the plates for 15 min while shaking. Afterward, absorbance is recorded at 550 and 620 nm using a microplate reader.

Cell Assay:^{^[1]}

Cell lines
Human leukemic Jurkat T cells and human leukemic monocytes THP-1 cells
Concentrations
10 to 82 μM (Jurkat cells); 20 to 163 μM (THP-1 cells)
Incubation Time
72 h
Method
The influence of TUPA on the cell viability is measured using the MTT assay. Cells are seeded in flat bottom 96-well plates at a density of 4 × 10⁵ cells/mL. Afterward, Jurkat cells are exposed to TUPA in concentrations ranging from 10 to 82 μM. THP-1 cells (4 × 10⁵ cells/mL) are exposed to concentrations ranging from 20 to 163 μM. After incubation for 72 h under humidified conditions (5% CO2, 37℃), 10 μL of MTT (5 mg/mL) per well are added and cells are further incubated for 3 h. Subsequently, 200 μL DMSO are added for cell lysis and formazan solubilization following an incubation of the plates for 15 min while shaking. Afterward, absorbance is recorded at 550 and 620 nm using a microplate reader.

References

[1] Zwicker P, et al. Proteomics. 2016, 16(23):2997-3008.

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