DNMT3A Assay Kit is use for measuring the amount of DNMT3A.
Epigenetic inactivation of genes plays a critical role in many important human diseases, especially in cancer. A core mechanism for epigenetic inactivation of the genes is methylation of CpG islands in genome DNA. Methylation of CpG islands involves the course in which DNA methyltransferases (Dnmts) transfer a methyl group from S-adenosyl-L-methionine to the fifth carbon position of the cytosines. At least three families of Dnmts have been identified in mammals now: Dnmt1, Dnmt2, and Dnmt3. The Dnmt3 family comprises of three different proteins: Dnmt3A, Dnmt3B, and Dnmt3L. Dnmt3A and Dnmt3B have been demonstrated to methylate both unmethylated and hemimethylated DNA equally and supposed to mediate de novo methylation together with Dnmt1. Increased activation or amount of Dnmt3 is believed to be involved in carcinogenesis, and other genetic and epigenetic diseases. Several methods such as Western blot are
For measuring Dnmt amount quantitatively from fresh tissue and cultured cells of human and mouse.
For research use only (RUO)
Store DA3 and DA5 at –20°C away from light. Store DA4, DA6 and 8 well assay strips at 4°C away from light.Store other components at room temperature. All components are stable for 6 months from date of shipment in proper storage.
DA1 (10X wash buffer) 25 mlDA2 (assay buffer) 3 mlDA3 (Dnmt3A standard, 20 µg/ml)* 20 µlDA4 (affinity antibody 100 µg/ml)* 50 µlDA5 (detection antibody 200 µg/ml)* 20 µlDA6 (developing solution) 12 mlDA7 (stop solution) 6 ml8 well substrate-coated strip (with frame) 12* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.
Features & Benefits:
Extremely fast procedure, which can be completed within 3 hours.Innovative colorimetric assay, which quantitatively measures Dnmt3A amount with no need for electrophoresis.Strip microplate format makes the assay flexible: manual or high throughput analysis.Simple, reliable, and consistent assay conditions.