MBD6 Antibody

Introduction to MBD6 Antibody

MBD6 (Methyl-CpG Binding Domain Protein 6) antibodies are specialized reagents designed to detect and study the MBD6 protein, a member of the methyl-CpG-binding domain family. Unlike other MBD proteins, MBD6 does not exhibit methyl-CpG binding activity in vitro but plays critical roles in chromatin remodeling, stem cell regulation, and cancer biology . Antibodies targeting MBD6 enable researchers to investigate its interactions with epigenetic complexes, such as the BAP1/ASXL complex, and its regulatory effects on stemness and differentiation .

Development and Validation of MBD6 Antibodies

MBD6 antibodies are generated using immunogens derived from conserved regions of the human MBD6 protein (UniProt ID: Q96DN6). Key validation steps include:

• Specificity Testing: Western blot and immunoprecipitation in MBD6 knockout cells to confirm absence of cross-reactivity .

• Epitope Mapping: Polyclonal antibodies often target the C-terminal region, which is critical for interactions with ASXL1-3 PHD domains .

• Functional Assays: Chromatin immunoprecipitation (ChIP-seq) confirms chromatin localization, particularly at regions enriched for H3K4me1, H3K27Ac, and other active histone marks .

Role in Epigenetic Complex Stabilization

MBD6 stabilizes the BAP1/ASXL complex by interacting with ASXL1-3 PHD fingers, enhancing BAP1’s deubiquitinase activity. Knockout studies show reduced BAP1 protein levels and increased H2AK119Ub marks .

Stemness and Differentiation Regulation

• Oct4 Interaction: MBD6 is a direct target of Oct4 and regulates stemness-related genes (e.g., REST, CDKs) in human adipose tissue-derived stem cells (hATSCs) .

• Differentiation Control: MBD6 promotes neural/endodermal differentiation while suppressing mesodermal lineages .

Chromatin Localization

Despite lacking methyl-DNA binding in vitro, MBD6 localizes to pericentric heterochromatin in 25% of cells, dependent on its MBD domain .

Applications in Biomedical Research

MBD6 antibodies are utilized in:

• Western Blot: Detecting endogenous MBD6 (~70 kDa) in HEK293T, SCLC, and stem cell lines .

• Immunoprecipitation: Identifying MBD6 interaction partners (e.g., BAP1, ASXL1-3) .

• ChIP-seq: Mapping MBD6-bound genomic regions enriched at active enhancers .

• Immunohistochemistry: Visualizing MBD6 expression in cancer tissues, particularly small cell lung cancer .

Challenges and Limitations

• DNA Binding Paradox: MBD6 does not bind methylated DNA in vitro despite its MBD domain, suggesting chromatin recruitment via protein interactions (e.g., ASXL PHD fingers) .

• Cell-Type Specificity: MBD6 expression varies significantly across tissues, with highest levels in testes and oocytes .