SpeedRun One-Hour MBD1 ELISA Kit (Colorimetric)

The SpeedRun™ One-Hour MBD1 ELISA Kit (Colorimetric) is a complete set of optimized reagents designed for measuring the amount of MBD1 using nuclear extracts or purified MBD1 proteins. The kit can be used with samples from a broad range of species, including human, mouse, and rabbit, and is suitable for cultured cells and fresh or 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 MBD1 proteins
• Sensitive: Limit of detection can be as low as 1 ng of MBD1 proteins
• Convenient: Includes MBD1 protein standard for quantification
• Flexible: Allows manual or high-throughput analysis with strip microplate format

Background Information
MBD1 (methyl-CpG-binding domain protein 1) is a member of the MBD protein family. MBD1 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, MBD1 is a novel target that has been accredited for cancer therapy. Some evidence has also linked this protein to neurodevelopment. The binding of MBD1 to methylated DNA may be affected by MBD1 mutation and biochemical or pharmacological intervention. Selective and potent MBD1 inhibition can induce gene re-expression, as seen with genetic disruption of MBD1 in cancer cells. Thus, detecting level of MBD1 is important in elucidating mechanisms of epigenetic regulation of gene activation and silencing, as well as benefiting cancer and neuro disease therapeutics.

Principle & Procedure
The kit contains all reagents necessary for the quantification of MBD1 levels. In this assay, MBD1 in the extracts is bound to strip-wells with a high binding buffer and is detected using MBD1 Detection Complex and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer. The MBD1 level is proportional to the OD intensity measured.

Starting Materials
Input materials can be nuclear extracts or purified MBD1 proteins. The amount of nuclear extracts for each assay can be 1 µg to 2 µg. The amount of purified proteins can be 1 ng to 50 ng with an optimal range of 10-20 ng, depending on the purity of the proteins.