MAML2 Antibody

What is MAML2 and what are its key biological functions?

MAML2 (mastermind like transcriptional coactivator 2) is a nuclear protein that functions as a transcriptional coactivator for NOTCH proteins. In humans, the canonical protein consists of 1156 amino acid residues with a molecular mass of approximately 125.2 kDa . It is widely expressed throughout the body, with particularly high expression levels detected in the placenta, salivary gland, and skeletal muscle . As a member of the Mastermind protein family, MAML2 plays a critical role in the Notch signaling pathway, which regulates various cellular processes including differentiation, proliferation, and apoptosis. MAML2 forms complexes with the intracellular domain of Notch (ICN) and the transcription factor CSL (RBP-Jκ) to regulate Notch target gene expression .

How do MAML2 antibodies differ in terms of specificity and cross-reactivity?

High-quality MAML2 antibodies demonstrate specificity for the target protein without cross-reactivity to other MAML family members. For instance, certain commercial antibodies specifically detect endogenous levels of total MAML2 protein without cross-reacting with MAML1 and MAML3 . This specificity is crucial for experimental validity, particularly in studies examining differential expression or function of MAML family members. Researchers should verify the specificity claims through validation studies before implementing MAML2 antibodies in their experimental protocols.

What species reactivity is typically available for MAML2 antibodies?

MAML2 antibodies commonly exhibit reactivity against human samples, with some antibodies also demonstrating cross-reactivity with mouse, rat, and monkey orthologs . The available search results indicate that many commercial antibodies primarily target human MAML2, though researchers studying animal models should carefully select antibodies with documented reactivity to their species of interest. For comparative studies across species, it is advisable to select antibodies that recognize conserved epitopes with demonstrated cross-species reactivity.

What are the primary applications for MAML2 antibodies in research?

MAML2 antibodies support multiple experimental applications, with Western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF) being the most commonly utilized techniques . Additional applications include flow cytometry (FCM), enzyme-linked immunosorbent assay (ELISA), and immunoprecipitation (IP) . The methodological approach varies by application:

• Western Blotting: Typically performed at dilutions of 1:1000, detecting MAML2 at approximately 160 kDa

• Immunohistochemistry: Performed on formalin-fixed tissues, often enhanced by epitope retrieval through boiling tissue sections in 10 mM Tris with 1 mM EDTA, pH 9.0 for 10-20 minutes

• Immunofluorescence: Generally employed at concentrations of 0.5-1 μg/mL

• Flow Cytometry: Utilized at 0.5-1 μg per million cells

• Immunoprecipitation: Performed at approximately 1:50 dilution

What positive controls are recommended for MAML2 antibody validation?

Based on published recommendations, MCF-7 cells serve as effective positive controls for MAML2 antibody validation in cellular systems . For tissue-based applications, pancreas, placenta, bladder carcinoma, and colon carcinoma samples are recommended as positive controls . When establishing a new MAML2 antibody in your laboratory, validation with these controls helps ensure antibody functionality and specificity prior to experimental use with test samples.

What are the optimal sample preparation methods for MAML2 immunohistochemistry?

For immunohistochemical applications, formalin-fixed, paraffin-embedded (FFPE) tissue sections represent the standard specimen format. Heat-induced epitope retrieval significantly enhances MAML2 detection in FFPE tissues. The recommended protocol involves boiling tissue sections in 10 mM Tris with 1 mM EDTA, pH 9.0 for 10-20 minutes followed by cooling at room temperature for 20 minutes . This retrieval step is crucial as formalin fixation can mask epitopes through protein cross-linking. Antibody concentrations between 0.5-1.0 μg/mL applied for 30 minutes at room temperature typically yield optimal results .

What is the diagnostic significance of MAML2 rearrangements in mucoepidermoid carcinoma?

MAML2 rearrangements serve as important diagnostic biomarkers in mucoepidermoid carcinoma (MEC), particularly when distinguishing MEC from the more aggressive adenosquamous carcinoma (ASC) . Studies indicate that MAML2 rearrangements have a sensitivity of approximately 60% for MEC diagnosis . The presence of MAML2 translocations provides objective confirmation of MEC diagnosis in challenging cases. In clinical practice, MAML2 testing has demonstrated significant impact on diagnostic decision-making, changing the working diagnosis in approximately 24% of cases in one comprehensive study .

What is the added value of MAML2 testing in the diagnostic workflow for salivary gland tumors?

MAML2 testing offers substantial clinical value in the diagnostic workflow for salivary gland tumors. Analysis of real-world testing practices demonstrates that MAML2 testing provides benefit in at least 38% of cases, or up to 81% when including confirmatory negative results in ASC cases . In practical terms, molecular testing for MAML2 has added diagnostic value in at least 1 in every 2.6 cases tested. Expert surveys further support this clinical utility, with confirmatory MAML2 testing in suspected MEC receiving a relative importance index of 0.8 on a standardized scale .

What factors should researchers consider when selecting between monoclonal and polyclonal MAML2 antibodies?

Researchers should consider several factors when choosing between monoclonal and polyclonal MAML2 antibodies:

For applications requiring high specificity and reproducibility, such as diagnostic immunohistochemistry, monoclonal antibodies like MAML2/1302 or 4A1 clones may be preferable . For applications prioritizing sensitivity, such as detection of low-abundance targets, polyclonal antibodies might offer advantages. The specific research question and intended application should guide antibody selection.

How do different MAML2 antibody clones compare in terms of epitope targeting?

Commercial MAML2 antibodies target various epitopes across the protein, each with potential advantages for specific applications:

• Antibodies targeting amino acids 347-506 are available as polyclonal reagents suitable for WB, IHC, and ELISA applications

• Monoclonal antibody clone 4A1 targets amino acids 796-894, optimized for WB and ELISA applications

• Antibodies directed against amino acids 780-814 are available in polyclonal format for WB and ELISA applications

• The MAML2/1302 clone is a versatile monoclonal antibody suitable for multiple applications including WB, FCM, IF, and IHC

Epitope selection affects antibody performance in different applications and under varying sample preparation conditions. For example, epitopes in highly conserved regions may increase cross-species reactivity, while epitopes in unique regions enhance specificity.

What quality control measures ensure reliable MAML2 antibody performance?

Rigorous quality control for MAML2 antibodies should include:

• Verification of protein specificity through Western blotting against recombinant MAML2 and related family members

• Cross-reactivity testing against MAML1 and MAML3 to confirm specificity

• Testing across multiple applications (WB, IHC, IF) using established positive controls

• Lot-to-lot consistency assessment to ensure reproducible performance

• Application-specific validation in relevant biological contexts (e.g., known MAML2-expressing tissues)

Researchers should review manufacturer validation data and, ideally, conduct independent validation before employing MAML2 antibodies in critical experiments.

How can MAML2 antibodies be employed in studies of Notch signaling pathway?

MAML2 antibodies serve as valuable tools for studying Notch signaling dynamics. MAML2 forms complexes with the intracellular domain of Notch (ICN) and CSL (RBP-Jκ) transcription factors to regulate Notch target gene expression . Researchers can utilize MAML2 antibodies for:

• Co-immunoprecipitation studies to identify protein-protein interactions within the Notch signaling complex

• Chromatin immunoprecipitation (ChIP) experiments to map MAML2 binding sites at Notch-responsive promoters

• Immunofluorescence co-localization studies to visualize MAML2 interactions with other Notch pathway components

• Proximity ligation assays to detect in situ protein interactions between MAML2 and Notch pathway members

These approaches contribute to understanding the molecular mechanisms underlying MAML2 function in normal development and disease contexts.

What is known about the relationship between MAML2 and MECT1/TORC1 in mucoepidermoid carcinomas?

MAML2 is frequently found fused with Mucoepidermoid carcinoma translocated gene 1 (MECT1, also known as WAMTP1 or TORC1) in patients with mucoepidermoid carcinomas and Warthin's tumors . This fusion event represents a defining molecular characteristic of these tumor types. Antibodies targeting different domains of MAML2 can be strategically employed to distinguish between wild-type MAML2 and fusion proteins. For instance, antibodies targeting the N-terminal region would detect both wild-type and fusion proteins, while those targeting regions beyond the fusion breakpoint would detect only wild-type MAML2. This differential detection capability enables researchers to study the specific contributions of fusion versus wild-type proteins in tumor biology.

How does MAML2 testing compare to other molecular diagnostic approaches in salivary gland tumors?

MAML2 testing represents one component of a comprehensive molecular diagnostic approach to salivary gland tumors. In a comparative assessment of diagnostic methodologies, MAML2 rearrangement testing demonstrates high specificity but moderate sensitivity (approximately 60%) for mucoepidermoid carcinoma . When integrated into diagnostic algorithms, MAML2 testing provides significant added value, with expert consensus supporting its utility particularly in distinguishing MEC from adenosquamous carcinoma .

The diagnostic power of MAML2 testing derives from its role as a composite biomarker - functioning as both an imperfect confirmation test for MEC and a highly specific exclusion tool for the diagnosis of ASC . This dual utility positions MAML2 testing as a valuable component of multimodal diagnostic approaches in salivary gland tumor classification.