MIER3 Antibody

Introduction to MIER3 Antibody

MIER3 antibodies are immunological reagents specifically designed to detect and visualize the MIER3 protein in various experimental contexts. These antibodies have become increasingly important in cancer research due to the growing recognition of MIER3's role as a potential tumor suppressor gene . As research tools, these antibodies enable scientists to study MIER3 expression patterns, protein interactions, and functional characteristics across different tissues and experimental conditions.

The development of specific and sensitive MIER3 antibodies has accelerated research into the biological functions of this protein, particularly in the context of cancer progression and metastasis. Multiple commercial suppliers now offer MIER3 antibodies with various specifications and applications, making them accessible for diverse research purposes in molecular biology, cell biology, and oncology.

Molecular Properties

MIER3 antibodies are predominantly produced in rabbits and available as polyclonal antibodies that recognize multiple epitopes on the MIER3 protein. These antibodies typically target specific amino acid sequences within the MIER3 protein structure. For example, some commercially available antibodies are developed against a recombinant protein corresponding to the amino acid sequence: VPESFMNEVSVNNLGVDFENHTHHITSAKMAVSVADFGSLSANETNGFISAHALHQHAALHSE .

The MIER3 protein itself has a calculated molecular weight of approximately 59-61 kDa, although the observed molecular weight in experimental conditions is often around 55 kDa . This discrepancy between calculated and observed molecular weights is not uncommon for proteins and may reflect post-translational modifications or protein folding characteristics.

Application Methods

MIER3 antibodies have been validated for multiple experimental applications, with Western blot (WB) being the most common. The recommended dilutions vary by supplier and application:

For immunohistochemistry applications on paraffin-embedded tissues, heat-induced epitope retrieval (HIER) at pH 6 is typically recommended . For immunocytochemistry, paraformaldehyde fixation followed by Triton X-100 permeabilization has been successfully employed .

Structure and Function

MIER3 (mesoderm induction early response 1, family member 3) belongs to the MIER protein family, which also includes MIER1 and MIER2. These proteins are fibroblast growth factor (FGF)-regulated immediate-early proteins that may have pivotal roles in FGF-regulated cellular activities .

The human MIER3 protein consists of approximately 550 amino acid residues with a calculated molecular mass of 61.4 kDa. Up to five different isoforms have been reported for this protein . Like other MIER family members, MIER3 functions primarily as a transcriptional repressor, localizing to the nucleus to regulate gene expression .

Cellular Localization and Expression

Expression analysis has revealed that MIER3 is downregulated in several cancer types compared to adjacent normal tissues. For instance, both mRNA and protein levels of MIER3 are significantly reduced in non-small cell lung cancer (NSCLC) tissues and cell lines compared to normal lung epithelial cells . Similarly, MIER3 expression is decreased in colorectal cancer tissues compared to normal colorectal tissues .

Role in Cancer Research

MIER3 antibodies have been instrumental in elucidating the role of MIER3 in cancer development and progression. Using these antibodies, researchers have demonstrated altered expression patterns of MIER3 in various cancer types, including non-small cell lung cancer and colorectal cancer .

Western blot and immunohistochemical analyses using MIER3 antibodies have shown that MIER3 is significantly downregulated in NSCLC tissues compared to adjacent normal tissues . This observation, combined with functional studies, has led to the identification of MIER3 as a potential tumor suppressor.

MIER3 as a Tumor Suppressor

Research utilizing MIER3 antibodies has provided substantial evidence for the tumor suppressor function of MIER3. In NSCLC, overexpression of MIER3 significantly inhibits cell proliferation, migration, and invasion, while promoting apoptosis and inducing cell cycle arrest . These effects have been verified both in vitro using cell culture models and in vivo using xenograft mouse models.

Similarly, in colorectal cancer, MIER3 overexpression has been shown to inhibit cell proliferation, migration, and invasion, as well as suppress tumor growth and metastasis in vivo . These findings collectively indicate that MIER3 functions as a tumor suppressor in multiple cancer types.

Signaling Pathway Research

MIER3 antibodies have facilitated investigations into the molecular mechanisms underlying MIER3's tumor-suppressive effects. In NSCLC, MIER3 has been shown to downregulate the Wnt/β-catenin signaling pathway, a key pathway involved in cancer development and progression .

Furthermore, co-immunoprecipitation assays using MIER3 antibodies have demonstrated that MIER3 can directly bind to histone acetyltransferase (HAT) p300, thereby inhibiting its activity . This interaction results in decreased acetylation of Histone3 and Histone4, which may contribute to transcriptional repression of genes involved in cancer progression.

In colorectal cancer, MIER3 inhibits cell proliferation and invasion partially through reduction of Sp1 and subsequent suppression of epithelial-mesenchymal transition (EMT) , a critical process in cancer metastasis.

Western Blot Analysis

Western blot analysis using MIER3 antibodies typically reveals a band of approximately 55 kDa, corresponding to the MIER3 protein . This technique has been widely used to compare MIER3 expression levels between normal and cancerous tissues, as well as among different cell lines.

For optimal results, most suppliers recommend antibody dilutions ranging from 1:500 to 1:2000 . Sample preparation typically involves protein extraction from tissues or cells, followed by denaturation, SDS-PAGE separation, and transfer to a membrane for immunodetection.

Immunohistochemistry and Immunofluorescence

Immunohistochemistry and immunofluorescence using MIER3 antibodies enable visualization of MIER3 protein distribution within tissues and cells. These techniques have revealed that MIER3 primarily localizes to the nucleus in most cell types, consistent with its role in transcriptional regulation .

For paraffin-embedded tissue sections, heat-induced epitope retrieval at pH 6 is typically recommended, followed by incubation with MIER3 antibody at dilutions ranging from 1:1000 to 1:2500 . For immunofluorescence on cultured cells, antibody concentrations of 0.25-2 μg/mL are commonly used .

ELISA Applications

Some MIER3 antibodies have been validated for enzyme-linked immunosorbent assay (ELISA) applications . This technique allows for quantitative measurement of MIER3 protein levels in biological samples, providing a complementary approach to Western blot analysis.

While specific protocols may vary, ELISA applications typically involve coating a plate with an antigen or capture antibody, followed by detection with a MIER3 antibody and a suitable detection system. The specific dilutions and conditions need to be optimized for each application and antibody.

Future Perspectives in MIER3 Research

The continued development and refinement of MIER3 antibodies will likely facilitate further investigations into the biological functions and clinical significance of MIER3. Several promising research directions include:

1. Exploration of MIER3's role in additional cancer types beyond NSCLC and colorectal cancer

2. Further elucidation of the molecular mechanisms underlying MIER3's tumor-suppressive effects

3. Investigation of MIER3 as a potential biomarker for cancer diagnosis or prognosis

4. Examination of MIER3's interactions with other proteins and signaling pathways

5. Development of therapeutic strategies targeting MIER3 or its downstream effectors

The availability of highly specific and sensitive MIER3 antibodies will be crucial for advancing these research areas and potentially translating findings into clinical applications.