CELSR3 Antibody, Biotin conjugated

What is CELSR3 and what are its structural characteristics?

CELSR3 (Cadherin EGF LAG seven-pass G-type receptor 3) belongs to the family of adhesion G protein-coupled receptors with a large extracellular region. This protein features a complex structure comprising nine cadherin domains, eight EGF repeats, and three laminin domains . It is also known by several aliases including Cadherin family member 11, Epidermal growth factor-like protein 1, Flamingo homolog 1 (hFmi1), Multiple epidermal growth factor-like domains protein 2, and is identified by UniprotID Q9NYQ7 .

CELSR3 is abundantly expressed in the adult brain along with its homolog CELSR2, where it plays crucial roles in neural development and function . From a functional perspective, CELSR3 is involved in planar cell polarity signaling pathways and forms specific complexes with other proteins such as Frizzled3 and Vangl2 on the plasma membrane, mediating intercellular communication essential for proper tissue organization .

What are the key specifications of commercially available CELSR3 Antibody, Biotin conjugated?

The commercially available CELSR3 Antibody, Biotin conjugated is a polyclonal antibody raised in rabbits using recombinant Human Cadherin EGF LAG seven-pass G-type receptor 3 protein (specifically amino acids 514-718) as the immunogen . This antibody preparation has the following technical specifications:

This antibody targets human CELSR3 and is particularly useful for signal transduction research applications .

How do biotinylated antibodies compare with other conjugates like PE for CELSR3 detection?

Biotinylated and PE-conjugated CELSR3 antibodies offer different advantages depending on the experimental context. The CELSR3 Antibody, Biotin conjugated is a polyclonal rabbit antibody primarily validated for ELISA applications . In contrast, the PE-conjugated variant (such as the Human CELSR3 PE-conjugated Antibody, FAB7278P) is a monoclonal mouse IgG, specifically clone #763103, optimized for flow cytometry applications .

Biotinylated antibodies provide versatility through the strong biotin-streptavidin interaction system, allowing for signal amplification and multi-step detection protocols. This makes them particularly useful in experiments requiring enhanced sensitivity or complex visualization schemes. The ability to pair with various streptavidin-conjugated detection reagents provides flexibility in experimental design.

PE-conjugated antibodies, with their direct fluorophore attachment (excitation = 488 nm, emission = 565-605 nm), are more suitable for direct detection methods like flow cytometry . They eliminate additional detection steps, reducing background and non-specific binding issues that can occur in multi-step protocols.

The choice between these conjugates should be guided by the specific experimental requirements, detection method, and whether signal amplification or direct detection is preferable.

What mechanisms underlie the interaction between CELSR3 and β-amyloid oligomers in neurodegenerative research?

Research has revealed that CELSR3 serves as a direct binding target for β-amyloid (Aβ) oligomers, with significant implications for neurodegenerative disease research . Studies using biotin-Aβ42 oligomers have demonstrated specific binding to CELSR3 but not to other tested proteins such as Vangl2 or Frizzled3, with an apparent dissociation constant (Kd) of approximately 40 nM equivalent of total Aβ peptide .

Domain-specific analyses have identified that Aβ oligomers bind specifically to the EGF repeats and laminin domains of CELSR3, not to its cadherin domains . This specificity is important as these same domains are partially involved in protein-protein interactions among planar cell polarity components. For instance, deletion of all eight EGF repeats and three laminin domains caused a 68% reduction in the interaction between CELSR3 and Frizzled3, while deletion of laminin G1 alone led to a 66% reduction .

The significance of this interaction lies in the mechanism through which Aβ oligomers may cause glutamatergic synapse degeneration by targeting planar cell polarity components in synapses. Researchers investigating neurodegenerative diseases can use CELSR3 Antibody, Biotin conjugated to:

• Track the binding interactions between Aβ oligomers and CELSR3

• Investigate how this binding affects CELSR3's normal function in planar cell polarity signaling

• Explore potential therapeutic interventions targeting this specific interaction

How can CELSR3 Antibody be used to investigate synaptic development and maintenance?

Knockout studies have demonstrated that CELSR3 plays a critical role in synaptic development and maintenance. When both CELSR2 and CELSR3 were knocked out in the CA1 region of the adult hippocampus using CRISPR-Cas9, researchers observed significant reductions in glutamatergic synapse numbers in the stratum radiatum . This was confirmed through multiple methodological approaches:

• Costaining with synaptic markers and quantification using the "synapse counter" Image J plugin, which measures puncta that are pre-synaptic, post-synaptic, or colocalized

• Sparse labeling of dendrites with AAV–human synapsin (hSyn)–mCherry to visualize dendritic spines, which showed significant reduction in spine numbers of CA1 pyramidal neurons following CELSR2 and CELSR3 knockout

Researchers can utilize CELSR3 Antibody, Biotin conjugated to:

• Visualize the localization of CELSR3 at synaptic junctions

• Track changes in CELSR3 expression during synapse formation and maintenance

• Investigate the differential distribution of CELSR3 in various neuronal populations

• Assess the impact of disease-associated mutations or treatments on CELSR3 expression and localization

This research direction is particularly valuable for understanding synaptopathies, which are increasingly recognized as central to many neurological and psychiatric disorders.

What methodological considerations are important when using biotinylated antibodies in systems with endogenous biotin?

When using CELSR3 Antibody, Biotin conjugated in experimental systems, researchers must consider the potential impact of endogenous biotin and biotin supplementation on their results. Studies have shown that biotin supplementation can significantly alter cellular function, including gene expression patterns .

Biotin affects the nuclear abundance of transcription factors Sp1 and Sp3, which act as transcriptional activators or repressors depending on the context . This can lead to downstream effects on protein expression and function. For example, biotin supplementation has been shown to decrease the expression of SERCA3 (sarco/endoplasmic reticulum Ca2+-ATPase 3) by more than 80% .

Researchers should implement the following methodological controls when using biotinylated antibodies:

• Include biotin blocking steps in protocols when working with samples exposed to varying biotin concentrations

• Consider using biotin-free media for cell culture experiments when possible

• Include controls to account for the potential effects of biotin on gene expression in the experimental system

• Validate findings with non-biotinylated detection methods when feasible

Additionally, researchers should be aware that biotin supplementation may induce cell stress by impairing protein folding in the endoplasmic reticulum . This stress response includes:

• Decreased sequestration of Ca2+ in the ER (14-24% reduction)

• Decreased secretion of proteins into extracellular space (e.g., 75% reduction in interleukin-2 secretion)

• Increased nuclear abundance of stress-induced transcription factors

• Increased abundance of stress-related proteins such as UBE1, GADD153, XBP1, and phosphorylated eIF2α

How can domain-specific analysis of CELSR3 improve understanding of its functional interactions?

Domain-specific analysis of CELSR3 has revealed important insights into its functional interactions with other proteins. Deletion studies have demonstrated that specific domains of CELSR3 play crucial roles in its binding interactions . For example:

Researchers can use CELSR3 Antibody, Biotin conjugated in combination with domain-specific constructs to:

• Map the binding sites between CELSR3 and its interaction partners

• Identify which domains are essential for specific cellular functions

• Develop domain-targeted therapeutics that could modulate specific CELSR3 interactions

• Investigate how disease-associated mutations in specific domains affect CELSR3 function

This approach is particularly valuable for understanding the mechanisms through which CELSR3 contributes to planar cell polarity signaling and how disruption of these interactions may contribute to neurological disorders.

What are the implications of CELSR3's role in planar cell polarity for neuronal migration and axon guidance?

CELSR3's role in planar cell polarity (PCP) signaling has significant implications for neuronal migration and axon guidance. The protein forms complexes with Frizzled3 on the plasma membrane of one cell, and with Vangl2 on the plasma membrane of neighboring cells . These interactions are essential for coordinating cell polarity across tissues.

In the nervous system, this coordination is crucial for processes such as:

• Proper neuronal migration during development

• Axon guidance and pathfinding

• Dendritic spine formation and maintenance

• Synaptogenesis and synaptic plasticity

Researchers using CELSR3 Antibody, Biotin conjugated can investigate:

• The spatiotemporal distribution of CELSR3 during developmental processes

• How disruption of CELSR3 expression or function affects neuronal positioning and connectivity

• The relationship between CELSR3-mediated PCP signaling and other guidance cue systems

• The potential role of CELSR3 in regenerative processes following neural injury

Understanding these mechanisms has broad implications for developmental neurobiology, regenerative medicine, and therapeutic approaches for neurodevelopmental disorders characterized by abnormal neuronal positioning or connectivity.

What are the optimal storage and handling conditions for maintaining CELSR3 Antibody, Biotin conjugated activity?

To maintain the optimal activity of CELSR3 Antibody, Biotin conjugated, researchers should adhere to the following storage and handling recommendations:

• Upon receipt, store the antibody at -20°C or -80°C for long-term storage

• Avoid repeated freeze-thaw cycles as this can lead to protein denaturation and loss of activity

• If frequent use is anticipated, consider aliquoting the antibody into single-use volumes before freezing

• The antibody is provided in a buffer containing 0.03% Proclin 300, 50% Glycerol, and 0.01M PBS at pH 7.4, which helps maintain stability

• When handling the antibody, maintain sterile conditions and use nuclease-free materials

• For short-term storage during experiments, keep the antibody on ice and return to appropriate storage conditions as soon as possible

Proper storage and handling are essential for maintaining antibody specificity and sensitivity in experimental applications. Degradation due to improper storage can lead to increased background, reduced signal strength, and potentially misleading experimental results.

How can researchers validate CELSR3 Antibody, Biotin conjugated specificity in their experimental systems?

Validating antibody specificity is crucial for obtaining reliable results. For CELSR3 Antibody, Biotin conjugated, researchers should consider implementing the following validation strategies:

• Positive and negative controls:

  • Use cell lines or tissues known to express or lack CELSR3

  • Include CELSR3 knockout or knockdown samples as negative controls

• Peptide competition assays:

  • Pre-incubate the antibody with the immunogen peptide (recombinant Human CELSR3 protein, amino acids 514-718)

  • A specific antibody will show reduced or eliminated binding in the presence of the competing peptide

• Multiple detection methods:

  • Confirm results using alternative antibodies targeting different epitopes of CELSR3

  • Compare results from different experimental techniques (e.g., ELISA, Western blot, immunohistochemistry)

• Molecular weight verification:

  • Ensure detected bands or signals correspond to the expected molecular weight of CELSR3

  • Account for post-translational modifications that may alter the observed molecular weight

• Cross-reactivity assessment:

  • Test the antibody against closely related proteins like CELSR1 and CELSR2

  • Especially important when studying systems where multiple CELSR family members are expressed

Thorough validation ensures that experimental observations genuinely reflect CELSR3 biology rather than non-specific interactions or technical artifacts.

How might combining CELSR3 Antibody with emerging technologies enhance neurodegenerative disease research?

The integration of CELSR3 Antibody, Biotin conjugated with emerging technologies presents exciting opportunities for advancing neurodegenerative disease research:

• Single-cell analysis technologies:

  • Combining CELSR3 antibody detection with single-cell RNA sequencing to correlate protein expression with transcriptomic profiles

  • Investigating cell-type specific responses to Aβ oligomer-CELSR3 interactions

• Super-resolution microscopy:

  • Utilizing biotinylated antibodies with streptavidin-conjugated quantum dots or organic fluorophores for nanoscale visualization of CELSR3 distribution

  • Examining the co-localization of CELSR3 with other PCP components at synaptic structures

• Proximity labeling techniques:

  • Adapting biotin-based proximity labeling methods (BioID, APEX) to identify novel CELSR3 interaction partners

  • Mapping the dynamic CELSR3 interactome during synaptic development and in disease states

• CRISPR-based approaches:

  • Combining antibody detection with CRISPR-Cas9 genome editing to study domain-specific functions of CELSR3

  • Developing CRISPR activation/interference systems to modulate CELSR3 expression in specific neural populations

These technological integrations can provide deeper insights into how CELSR3 dysfunction contributes to neurodegenerative pathology and potentially identify novel therapeutic targets.

What therapeutic potential exists in targeting CELSR3-mediated pathways in neurological disorders?

Given CELSR3's role in glutamatergic synapse development and maintenance, and its specific interaction with Aβ oligomers, targeting CELSR3-mediated pathways holds significant therapeutic potential:

• Disrupting Aβ-CELSR3 interactions:

  • Developing peptide mimetics or small molecules that block the binding of Aβ oligomers to CELSR3's EGF repeats and laminin domains

  • This approach could potentially prevent Aβ-induced synapse degeneration while preserving normal CELSR3 function

• Modulating CELSR3 expression:

  • Investigating whether upregulating CELSR3 could compensate for synapse loss in neurodegenerative conditions

  • Exploring whether selective enhancement of CELSR3 in specific brain regions could promote synaptic maintenance

• Targeting downstream effectors:

  • Identifying and modulating the signaling pathways activated by CELSR3 that specifically contribute to synaptic stability

  • Developing compounds that mimic or enhance protective CELSR3 signaling pathways

• Biomarker development:

  • Utilizing CELSR3 Antibody, Biotin conjugated to detect soluble CELSR3 fragments or CELSR3-Aβ complexes that might serve as diagnostic or prognostic biomarkers

Research in this direction could lead to novel therapeutic strategies for neurodegenerative diseases characterized by synapse loss, particularly Alzheimer's disease where Aβ oligomers play a central pathogenic role.