DNM2 Antibody

Introduction to DNM2 Antibody

DNM2 antibodies are immunological reagents developed for the detection and study of dynamin 2 (DNM2), a protein encoded by the DNM2 gene. These antibodies have become essential tools in cellular and molecular biology research, particularly for investigating membrane dynamics, endocytosis, and cytoskeletal regulation. DNM2 antibodies bind specifically to epitopes on the dynamin 2 protein, enabling its detection in various experimental contexts .

The canonical human dynamin 2 protein has 870 amino acid residues with a molecular mass of approximately 98-101 kDa. Alternative splicing produces multiple isoforms, with the protein being ubiquitously expressed across many tissue types . DNM2 antibodies have been cited in over 70 research publications, highlighting their importance in scientific investigations .

Structure and Properties of DNM2

Before delving deeper into DNM2 antibodies, it's important to understand the target protein itself. Dynamin 2 is a large GTPase that plays critical roles in multiple cellular functions:

• Membrane trafficking and cytoskeleton regulation

• Clathrin-mediated endocytosis (CME)

• Exocytic processes and clathrin-coated vesicle formation from the trans-Golgi network

• PDGF-stimulated macropinocytosis

• Organization of the actin cytoskeleton

During vesicular trafficking, DNM2 associates with membranes through lipid binding and self-assembles into ring-like structures. Through oligomerization, it forms a helical polymer around vesicle membranes, facilitating vesicle scission . The protein contains several functional domains that enable these complex cellular activities, making it a fascinating subject for antibody-based research.

DNM2 Isoforms and Tissue Expression

DNM2 exists in multiple isoforms, including ubiquitous (Ub-DNM2) and muscle-specific (M-DNM2) variants. Research has shown that M-DNM2, which includes exon 12b, represents approximately 21% of all pan-DNM2 at embryonic day 18.5, increasing to 44% at 2 weeks and 49% at 7 weeks of age in mouse models. This corresponds with muscle hypertrophy and maturation of intracellular organization in myofibers .

The two most expressed DNM2 transcripts in adult mouse muscle are:

1. Ubiquitous DNM2 including exon 10a

2. Muscle-specific DNM2 including exon 10a

Types of DNM2 Antibodies

Various types of DNM2 antibodies are commercially available, each with specific characteristics suitable for different research applications.

Based on Production Method

Based on Host Species

DNM2 antibodies are produced in different host species, most commonly:

1. Mouse-derived antibodies - typically monoclonal, often used for human DNM2 detection

2. Rabbit-derived antibodies - both polyclonal and recombinant monoclonal versions available

Based on Conjugation

Many DNM2 antibodies are available in unconjugated forms, but conjugated versions offer additional utilities:

Applications of DNM2 Antibodies

DNM2 antibodies are utilized in various laboratory techniques to study dynamin 2 expression, localization, and function.

Western Blotting (WB)

Western blotting is one of the most common applications for DNM2 antibodies. These antibodies typically detect a band of approximately 98-101 kDa, corresponding to the dynamin 2 protein . Recommended dilutions vary by product:

• Mouse anti-Human DNM2 antibody detects a band of 101 kDa in K562 cell lysates

• DNM2 Rabbit Polyclonal antibody (SH-A11664): WB dilution 1:100-1000

• DNM2 Mouse Monoclonal (68209-1-Ig): WB dilution 1:5000-1:50000

Immunocytochemistry/Immunofluorescence (ICC/IF)

DNM2 antibodies are effective for visualizing the cellular localization of dynamin 2 protein. Recommended dilutions include:

• Rabbit polyclonal antibodies: IF dilution 1:100-1:500

• Mouse monoclonal antibodies: IF/ICC dilution 1:200-1:800

ELISA

Many DNM2 antibodies are validated for use in ELISA assays, with typical dilution ranges of 1:500-1:3000 for rabbit polyclonal antibodies .

Additional Applications

Some specialized DNM2 antibodies have been validated for flow cytometry and other techniques, expanding the research possibilities .

Cross-Reactivity

DNM2 antibodies demonstrate varying species cross-reactivity, which is important to consider when designing experiments:

DNM2 in Disease Pathophysiology

Research using DNM2 antibodies has contributed significantly to understanding disease mechanisms associated with dynamin 2 dysfunction.

Genetic Disorders

Heterozygous mutations in the DNM2 gene are associated with two tissue-specific diseases:

1. Charcot-Marie-Tooth disease (CMT) - a peripheral nerve disorder

2. Autosomal dominant centronuclear myopathy (CNM) - a muscle disease characterized by abnormal nuclei positioning

In a research study using the p.R465W mutation knock-in mouse model (Dnm2^RW/+), investigators demonstrated that reducing DNM2 levels could rescue pathophysiological features of CNM. This finding has important therapeutic implications for patients with DNM2-related disorders .

Cancer and Hematological Malignancies

DNM2 antibodies have been instrumental in studying the role of dynamin 2 in cancer development and progression. Research has shown:

• DNM2 is more highly expressed in both B-cell and T-cell acute lymphoblastic leukemia (ALL) compared to normal cells

• High DNM2 expression in B-cell ALL is associated with:

  • White blood cell count ≥30 × 10^9/L (79% vs. 42%; P = 0.003)

  • Higher frequency of lymphadenopathy (61% vs. 23%; P = 0.002)

  • Briefer median relapse-free survival and 5-year survival

Research Findings on DNM2

DNM2 antibodies have facilitated numerous important discoveries about dynamin 2 regulation and function.

Transcriptional Regulation of DNM2

Research has revealed that the transcription factor Ikaros (encoded by IKZF1) directly binds to the DNM2 promoter region and suppresses its expression. This regulatory mechanism was demonstrated through:

• ChIP-seq data identifying Ikaros binding peaks in the DNM2 promoter region

• Luciferase reporter assays confirming Ikaros-mediated suppression of DNM2 promoter activity

• qPCR and western blot assays showing increased DNM2 expression following Ikaros knockdown

IKZF1 Deletion and DNM2 Expression

Studies have shown a strong correlation between IKZF1 deletion and DNM2 expression:

This correlation was confirmed in multivariate analyses (HR 6.151, 95% CI 1.401-27.000; P = 0.016) .

Therapeutic Targeting of DNM2

Research using DNM2 antibodies has identified potential therapeutic strategies targeting dynamin 2:

1. DNM2 reduction in genetic disorders: Both adeno-associated virus-shRNA and antisense oligonucleotides targeting DNM2 have successfully restored muscle mass, histopathology, and ultrastructural features in mouse models of centronuclear myopathy

2. DNM2 inhibition in leukemia: The DNM2 inhibitor MiTMAB demonstrated dose-dependent inhibition of leukemia cell proliferation. Furthermore, combining MiTMAB with CK2 inhibitors (TBB or CX-4945) showed synergistic effects in suppressing leukemia cell proliferation

Differential Roles of DNM2 Isoforms

Research using specific antibodies against the muscle-specific DNM2 isoform (containing exon 12b) has revealed distinct functional roles in skeletal muscle. Studies in Dnm2ex12b knockout mice showed:

• Normal muscle force production

• No obvious histological anomalies

• Normal sarcomere organization

• Increased misorientation of T-tubules

Production and Purification Methods

DNM2 antibodies are produced using various methods:

• Affinity chromatography on Protein G from tissue culture supernatant

• Antigen affinity purification

Immunogens Used

Different immunogens are used to generate DNM2 antibodies, affecting their specificity:

• Recombinant fusion proteins containing specific DNM2 sequences

• Synthetic peptides corresponding to regions within human DNM2

• DNM2 recombinant proteins

Future Directions in DNM2 Antibody Research

DNM2 antibody research continues to evolve, with several promising directions:

1. Development of isoform-specific antibodies: As research reveals distinct functions for different DNM2 splice variants, antibodies capable of distinguishing these isoforms become increasingly valuable

2. Therapeutic antibodies: Building on findings that DNM2 reduction can ameliorate disease symptoms, therapeutic antibodies or antibody-drug conjugates targeting DNM2 might be developed

3. Diagnostic applications: Given the association between DNM2 expression and disease prognosis in certain cancers, DNM2 antibodies could potentially be developed into diagnostic tools

4. Structural studies: Antibodies that recognize specific conformational states of dynamin 2 could provide insights into its mechanistic functions

The continued refinement and development of DNM2 antibodies will undoubtedly contribute to advancing our understanding of this important protein's roles in health and disease.