DIAPH2 Mouse mAb: Target Biology, Applications & Protocol

DIAPH2 Mouse mAb is a monoclonal antibody raised in mouse that specifically targets the human DIAPH2 (Diaphanous-related formin-2) protein, enabling reliable detection and functional study of this key actin cytoskeleton regulator across multiple experimental platforms.

Product Name

DIAPH2 Mouse mAb

SKU / Catalog Number

Ab349495

Host Species

Mouse

Antibody Type

Monoclonal Antibody (mAb)

Target Protein

DIAPH2 (Diaphanous-related formin-2)

Reference Clone Code

KC-40807 (absea)

Target Overview: What Is DIAPH2?

DIAPH2, also known as Diaphanous-related formin-2 (DAAM2 in some literature contexts, though DIAPH2 is a distinct gene), is a member of the diaphanous-related formin (DRF) family of proteins. Formins are well-characterized nucleators and elongators of unbranched actin filaments and play essential roles in regulating the actin cytoskeleton. DIAPH2 is involved in cytokinesis, cell polarity, filopodia formation, and microtubule dynamics, making it a multifunctional regulator of cell morphology and division.

DIAPH2 contains characteristic formin homology domains (FH1 and FH2) responsible for profilin binding and actin polymerization, as well as an autoinhibitory Diaphanous-inhibitory domain (DID) regulated by Rho GTPases. Dysregulation of DIAPH2 has been implicated in cancer progression, premature ovarian failure, and neurodevelopmental disorders, underscoring its biomedical relevance.

Product Description & Reagent Specifications

The DIAPH2 Mouse mAb (catalog number Ab349495) is a mouse monoclonal antibody generated against DIAPH2 protein. It is offered by Aladdin Scientific as a high-quality research reagent for detection of DIAPH2 in experimental samples. The reference clone designation from the originating manufacturer is KC-40807 (absea). Monoclonal antibodies offer superior lot-to-lot consistency and epitope specificity compared to polyclonal sera, making Ab349495 a reliable choice for reproducible research.

Note: Specific technical parameters including immunogen sequence, clonality confirmation, isotype, conjugation, recommended dilutions, validated species reactivity, and storage conditions should be verified against the current official datasheet available on the Aladdin Scientific product page or by contacting technical support, as these details were not provided in the source material.

Recommended Protocols & Application Guidelines

Mouse monoclonal antibodies targeting formin-family proteins such as DIAPH2 are routinely employed in the following assay formats. Always consult the official product datasheet for application-specific validation status and optimized conditions for Ab349495:

• Western Blot (WB): Denature and reduce cell or tissue lysates, resolve by SDS-PAGE, and transfer to PVDF or nitrocellulose membrane. Block with 5% non-fat milk or BSA in TBST. Incubate with primary antibody at the manufacturer-recommended dilution overnight at 4 °C. Use an appropriate HRP-conjugated anti-mouse secondary antibody for detection. The expected molecular weight of DIAPH2 should be confirmed against the current datasheet.
• Immunofluorescence (IF) / Immunocytochemistry (ICC): Fix cells with 4% paraformaldehyde, permeabilize with 0.1–0.3% Triton X-100, block, and incubate with the primary antibody. Visualize with a fluorophore-conjugated anti-mouse secondary antibody. Co-staining with phalloidin (F-actin marker) can provide informative context for DIAPH2 localization studies.
• Immunoprecipitation (IP): Lyse cells under non-denaturing conditions. Incubate lysate with Ab349495 and capture immune complexes using Protein A/G beads. Elute and analyze by SDS-PAGE/WB.
• Immunohistochemistry (IHC): Perform antigen retrieval on FFPE tissue sections (heat-induced, citrate buffer pH 6.0 is commonly used), block endogenous peroxidase, and apply primary antibody followed by an HRP-polymer anti-mouse detection system.

Optimal working dilutions, incubation times, and validated applications must be confirmed from the official Ab349495 datasheet.

Storage, Handling & Quality Assurance

To maintain antibody activity and ensure experimental reproducibility, follow general best practices for monoclonal antibody storage and handling:

• Storage: Store as directed on the product datasheet. Antibodies are typically supplied in a stabilizing buffer (e.g., PBS with BSA and sodium azide) and stored at –20 °C for long-term use or at 4 °C for short-term use. Avoid repeated freeze-thaw cycles by aliquoting upon first use.
• Handling: Thaw on ice, mix gently (do not vortex), and centrifuge briefly before use to collect any condensate. Work in a clean environment to prevent contamination.
• Quality: As a monoclonal antibody, Ab349495 offers high epitope specificity and batch-to-batch reproducibility. Each lot is subject to quality control testing by the manufacturer. Certificate of Analysis (CoA) data are available upon request from Aladdin Scientific.

Specific storage temperature, buffer composition, concentration, and shelf life must be confirmed from the official product datasheet or CoA.

Frequently asked questions

What is the catalog number for DIAPH2 Mouse mAb at Aladdin Scientific?

The catalog number for DIAPH2 Mouse mAb at Aladdin Scientific is Ab349495. The reference clone code from the originating manufacturer (absea) is KC-40807.

What applications is DIAPH2 Mouse mAb (Ab349495) validated for?

Common applications for mouse monoclonal antibodies against DIAPH2 include Western Blot (WB), Immunofluorescence (IF), Immunohistochemistry (IHC), and Immunoprecipitation (IP). However, the specific validated applications for Ab349495 should be confirmed by consulting the official product datasheet available from Aladdin Scientific or by contacting technical support, as this information was not fully specified in the available source material.

Why use a monoclonal antibody rather than a polyclonal antibody for DIAPH2 detection?

Monoclonal antibodies like Ab349495 recognize a single, defined epitope on the DIAPH2 protein, providing superior specificity, reduced background, and excellent lot-to-lot reproducibility. This makes them particularly advantageous for quantitative assays and studies requiring consistent, reproducible results across experiments and laboratories.

How should I store DIAPH2 Mouse mAb (Ab349495) to maintain its activity?

General best practices recommend storing monoclonal antibodies at –20 °C for long-term preservation and at 4 °C for short-term use. To avoid activity loss, aliquot the antibody upon first thaw to prevent repeated freeze-thaw cycles. Specific storage conditions, including buffer composition and recommended temperature, should be verified from the official Ab349495 product datasheet or Certificate of Analysis.

What is the biological significance of DIAPH2 as a research target?

DIAPH2 is a member of the diaphanous-related formin family and plays key roles in actin filament nucleation, cytokinesis, filopodia formation, and microtubule regulation. Its dysregulation has been associated with cancer progression, premature ovarian failure, and neurodevelopmental conditions, making it a target of significant interest in cell biology, oncology, and reproductive medicine research.

References

1. Aladdin Scientific product page: DIAPH2 Mouse mAb, Cat. No. Ab349495 (reference clone KC-40807, absea).
2. Chesarone MA, DuPage AG, Goode BL. Unleashing formins to remodel the actin and microtubule cytoskeletons. Nat Rev Mol Cell Biol. 2010;11(1):62-74. doi:10.1038/nrm2816
3. Rivero F, et al. The Diaphanous-related formins: an emerging subfamily of effectors of Rho GTPases. Cell Mol Life Sci. 2005;62(12):1385-1396.

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.