DRG1 Antibody, FITC conjugated

Introduction to DRG1 Antibody, FITC Conjugated

DRG1 Antibody, FITC conjugated is an immunological reagent created by linking fluorescein isothiocyanate (FITC) to antibodies that specifically recognize the Developmentally Regulated GTP Binding Protein 1 (DRG1). FITC is a commonly used fluorescent dye that emits green light when excited by appropriate wavelengths, allowing for visual detection through fluorescence microscopy or quantitative analysis through flow cytometry. The conjugation of FITC to DRG1-specific antibodies creates a powerful tool for directly detecting and visualizing DRG1 protein in experimental settings without requiring secondary detection methods .

The development of such conjugated antibodies has significantly advanced research capabilities in cellular and molecular biology, offering improved sensitivity and convenience over traditional indirect detection methods. These antibodies are particularly valuable in applications requiring direct visualization or quantification of protein expression, localization, and interactions in complex biological samples.

Structure and Expression

DRG1, also known as NDRG1 (N-myc downstream-regulated gene 1 protein), CAP43, RTP, or Protein NDRG1, is a highly conserved cytoplasmic protein that belongs to the NDR family of proteins . The protein contains multiple phosphorylation sites, with phosphorylation at threonine 346 (T346) being particularly important for its function and regulation . DRG1 is widely expressed in various human tissues and cells, with its expression levels often altered in pathological conditions, particularly in cancer.

Biological Functions

DRG1 functions as a stress-responsive protein involved in multiple cellular processes, including:

• Hormone responses and signal transduction

• Cell growth regulation and differentiation

• Tumor suppression in multiple cell types

• Cellular trafficking and vesicular recycling

• Maintenance and development of peripheral nerve myelin sheaths

• Regulation of p53-dependent apoptosis and cell cycle control

• Protection of cells from spindle disruption damage

• Regulation of microtubule dynamics and maintenance of euploidy

DRG1 plays a critical role in the vesicular recycling of proteins like CDH1 (E-cadherin) and TF (transferrin). It may also function in lipid trafficking and is necessary but not sufficient for p53-mediated caspase activation and apoptosis . These diverse functions highlight the protein's importance in normal cellular physiology and its potential role in disease pathogenesis.

Antibody Generation

DRG1 antibodies are typically generated using recombinant protein fragments or synthetic peptides corresponding to specific regions of the human DRG1 protein as immunogens. For instance, some commercially available DRG1 antibodies target amino acids 173-228 of the human DRG1 protein . These antibodies may be polyclonal, derived from rabbit immune serum, or monoclonal, produced from a single B cell clone, offering high specificity for the target epitope .

FITC Conjugation Process

The conjugation of FITC to DRG1 antibodies follows established protocols in immunochemistry. The process involves crosslinking the primary antibody with the FITC fluorophore through chemical reactions targeting the antibody's amino groups. This procedure is carefully controlled to maintain antibody specificity and functionality while achieving optimal fluorescent labeling . The resulting conjugate retains the binding specificity of the original antibody while gaining fluorescent properties that enable direct detection.

Flow Cytometry

One of the primary applications of DRG1 Antibody, FITC conjugated is flow cytometry. This technique allows for rapid quantification of DRG1 expression in cell populations and can identify specific cell subsets based on DRG1 expression patterns . The direct FITC conjugation eliminates the need for secondary antibodies, simplifying the experimental procedure and reducing background signal.

Immunofluorescence Microscopy

FITC-conjugated DRG1 antibodies are valuable tools for localizing and visualizing DRG1 protein within cells or tissues through immunofluorescence microscopy. This application provides insights into the subcellular distribution of DRG1 under various conditions and can reveal changes in localization in response to different stimuli or in disease states . The recommended dilution for immunofluorescence applications typically ranges from 1:50 to 1:200 .

Other Applications

Although less common, FITC-conjugated DRG1 antibodies may also be used in other applications such as:

• Immunohistochemistry (IHC) for tissue sections

• Western blotting for protein detection following electrophoretic separation

• ELISA-based detection systems

For these applications, the appropriate dilution must be empirically determined, although manufacturers often provide recommended starting dilutions.

Light Sensitivity

A critical consideration for FITC-conjugated antibodies is their sensitivity to light. Continuous exposure to light causes the FITC fluorophore to gradually lose its fluorescence, a phenomenon known as photobleaching . Therefore, these antibodies should be protected from light during storage and handling, and exposure time should be minimized during experimental procedures.

Working Solution Preparation

When preparing working solutions of DRG1 Antibody, FITC conjugated, it is recommended to dilute the antibody in appropriate buffers, such as Phosphate-Buffered Saline (PBS) containing protein stabilizers like bovine serum albumin (BSA) or fetal bovine serum (FBS) . The optimal dilution depends on the specific application and should be determined empirically, although manufacturers often provide recommended dilution ranges.

DRG1 in Cancer Research

DRG1 has been identified as a tumor suppressor in multiple cancer types, making DRG1 Antibody, FITC conjugated an important tool in cancer research. Studies have shown that DRG1 expression is downregulated in various human cancers, and its restoration can inhibit tumor growth and metastasis . The protein functions in the p53-dependent mitotic spindle checkpoint and protects cells from spindle disruption damage, suggesting a role in maintaining genomic stability .

DRG1 in Neurological Research

DRG1 plays a crucial role in the maintenance and development of the peripheral nerve myelin sheath, with mutations in the DRG1 gene associated with Charcot-Marie-Tooth disease, a hereditary motor and sensory neuropathy . FITC-conjugated DRG1 antibodies enable researchers to visualize DRG1 localization in neuronal cells and tissues, providing insights into its function in normal and pathological neural development.

DRG1 in Cell Biology

The multifunctional nature of DRG1 in cell trafficking, vesicular recycling, and cytoskeletal dynamics makes it a subject of interest in basic cell biology research. FITC-conjugated DRG1 antibodies facilitate the study of DRG1's interactions with other cellular components and its dynamic localization in response to various stimuli, contributing to our understanding of fundamental cellular processes .