SpeedRun One-Hour MBD2 ELISA Kit (Colorimetric)

The SpeedRun™ One-Hour MBD2 ELISA Kit (Colorimetric) is a complete set of optimized reagents designed for measuring the amount of MBD2 using nuclear extracts or purified MBD2 proteins from a broad range of species such as human, mouse and rabbit, in a variety of forms including, but not limited to, cultured cells and fresh/frozen tissues. The kit has the following advantages and features:

• Quick: Colorimetric assay completed within 1 hour
• Versatile: Works with nuclear extracts from cells/tissues or purified MBD2 proteins
• Sensitive: Limit of detection can be as low as 1 ng of MBD2 proteins
• Convenient: Includes MBD2 protein standard for quantification
• Flexible: Allows manual or high-throughput analysis with strip microplate format
  

Background Information
MBD2 (methyl-CpG-binding domain protein 2) is a member of the MBD protein family. MBD2 selectively binds to methylated DNA and suppresses transcription from a methylated target gene through recruiting transcriptional repressor complexes. As an epigenetic reader of methylated CpG DNA, MBD2 is a novel target that has been accredited for cancer therapy. Some evidence has also linked this protein to neurodevelopment. The binding of MBD2 to methylated DNA may be affected by MBD2 mutation and biochemical or pharmacological intervention. Selective and potent MBD2 inhibition can induce gene re-expression, as seen with genetic disruption of MBD2 in cancer cells. Thus, detecting level of MBD2 is important in elucidating mechanisms of epigenetic regulation of gene activation and silencing, as well as benefiting cancer and neuro disease therapeutics.

Principle & Procedure
This kit contains all reagents necessary for the quantification of MBD2 levels. In this assay, MBD2 in the extracts is bound to strip wells with a high binding buffer and is detected using the MBD2 Detection Complex and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer. The MBD2 level is proportional to the OD intensity measured.

Starting Materials
Input materials can be nuclear extracts or purified MBD2 proteins. The amount of nuclear extracts for each assay can range from 1 to 2 µg. The amount of purified proteins can range from 1 to 50 ng, with an optimal range of 10 to 20 ng, depending on the purity of the proteins.