C1QTNF3 Antibody

What is C1QTNF3 and why is it important in research?

C1QTNF3 (Complement C1q Tumor Necrosis Factor-Related Protein 3), also known as CTRP3 or CORS26, belongs to the highly conserved family of Acrp30/Adiponectin paralogs known as C1q and TNF-related protein family. This protein has gained significant research attention because it functions as an adipokine that regulates hepatic glucose output and may play roles in skeletal development . Unlike adiponectin, which is expressed exclusively by differentiated adipocytes, CTRP3 is expressed in a wide variety of tissues, making it important for understanding cross-tissue metabolic communication . Research has also demonstrated that CTRP3 stimulates testosterone production through the cAMP/PKA signaling pathway, suggesting its involvement in reproductive endocrinology .

What is the molecular structure of C1QTNF3?

C1QTNF3 shares a modular organization common to the C1q and TNF-related protein family. Its structure comprises an N-terminal signal peptide, a short variable region, a collagenous domain, and a C-terminal globular domain . C1qTNF proteins form trimeric structures that can assemble into hexameric and higher-order molecular forms, which is crucial for their biological function . The protein's structural features have evolutionary links to both TNF and C1q-containing proteins, suggesting these proteins arose from a common ancestral innate immunity gene . Multiple isoforms of human CTRP3 have been reported, adding complexity to functional studies of this protein .

How should C1QTNF3 antibodies be stored for optimal stability?

For optimal stability and activity retention, C1QTNF3 antibodies should be stored following specific protocols. The recommended storage conditions are:

It is strongly recommended to use a manual defrost freezer and avoid repeated freeze-thaw cycles as these can significantly reduce antibody activity . For long-term storage, aliquoting the reconstituted antibody is advisable to minimize damage from freeze-thaw cycles . Some antibody formulations are supplied in PBS with 0.02% sodium azide and 50% glycerol, which helps maintain stability .

What are the validated applications for C1QTNF3 antibodies?

C1QTNF3 antibodies have been validated for multiple experimental applications, allowing researchers to investigate this protein across various contexts. The table below summarizes the main applications with corresponding sample types:

When designing experiments, it's important to note that optimal dilutions should be determined by each laboratory for each specific application . For ELISA development, certain antibody pairs have been validated, such as the Mouse Anti-Human CTRP3/C1qTNF3/CORS26 Monoclonal Antibody functioning as a capture antibody when paired with a corresponding detection antibody .

How can I ensure specificity when using C1QTNF3 antibodies across species?

Cross-reactivity is an important consideration when working with C1QTNF3 antibodies across different species. Some mouse-derived antibodies show approximately 5% cross-reactivity with recombinant human CTRP3/C1qTNF3/CORS26 and less than 2% cross-reactivity with recombinant mouse Acrp30 . For studies involving multiple species, it's advisable to:

• Select antibodies validated for your species of interest. Some antibodies are specifically validated for human, mouse, and rat samples .

• Perform validation experiments using positive and negative controls from your species.

• Consider using antibodies raised against conserved epitopes when cross-species detection is desired.

• Be aware that human and mouse CORS26 share approximately 95% amino acid sequence homology, which explains the potential for cross-reactivity .

For critical studies, confirming specificity through knockout/knockdown validation or competitive blocking experiments is recommended to ensure the observed signals are specific to C1QTNF3.

What is the recommended ELISA setup for quantifying C1QTNF3?

For researchers developing ELISA assays to quantify C1QTNF3, a sandwich ELISA approach has been validated. The protocol involves:

• Coating a Clear Polystyrene Microplate with Mouse Anti-Human CTRP3/C1qTNF3/CORS26 Monoclonal Antibody as the capture antibody.

• Blocking non-specific binding sites.

• Adding samples containing CTRP3/C1qTNF3/CORS26 protein.

• Incubating with biotinylated Mouse Anti-Human CTRP3/C1qTNF3/CORS26 Monoclonal Antibody as the detection antibody.

• Detection using Streptavidin-HRP followed by appropriate substrate solution.

• Stopping the enzymatic reaction and measuring absorbance .

For standard curve preparation, recombinant Human CTRP3/C1qTNF3/CORS26 protein can be serially diluted 2-fold . Commercial ELISA development kits are available for convenience, such as the Human CTRP3/C1qTNF3/CORS26 DuoSet ELISA Kit .

How can C1QTNF3 antibodies be used to study adipokine signaling pathways?

C1QTNF3, as an adipokine, participates in metabolic signaling pathways that can be studied using specific antibodies. Research has shown that CTRP3 stimulates testosterone production through the cAMP/PKA signaling pathway . To investigate these signaling mechanisms:

• Use antibodies in Western blot analysis to detect changes in CTRP3 expression under different metabolic conditions.

• Combine with phospho-specific antibodies to track activation of downstream effectors in the cAMP/PKA pathway.

• Employ co-immunoprecipitation with C1QTNF3 antibodies to identify binding partners.

• Design immunofluorescence experiments to track subcellular localization changes in response to stimuli.

For comprehensive pathway analysis, consider combining antibody-based techniques with functional assays such as reporter gene assays or metabolic measurements. Previous studies have successfully used C1QTNF3 antibodies to demonstrate this protein's role in regulating hepatic glucose output, making it a valuable tool for metabolic research .

What considerations are important when analyzing oligomeric states of C1QTNF3?

C1QTNF3 forms trimeric structures that can assemble into hexameric and higher-order molecular forms, which is critical for its function . When studying these oligomeric states:

• Sample preparation is crucial – use non-reducing conditions when appropriate to preserve disulfide bonds that stabilize oligomers.

• Consider using native PAGE rather than SDS-PAGE to maintain protein-protein interactions.

• For Western blotting, be aware that different bands may represent different oligomeric states:

  • Monomers: ~26-30 kDa

  • Trimers: ~75-90 kDa

  • Higher oligomers: >150 kDa

Research has shown that cysteine-mediated oligomerizations are important for C1QTNF3 function . Employing size exclusion chromatography in combination with C1QTNF3 antibody detection can provide additional confirmation of oligomeric states in experimental samples.

How can I investigate tissue-specific expression patterns of C1QTNF3?

Unlike adiponectin, which is expressed exclusively in adipocytes, C1QTNF3 shows expression across multiple tissues . To study tissue-specific expression:

• Use immunohistochemistry (IHC-P) with C1QTNF3 antibodies at recommended dilutions (e.g., 1/20-1/200) .

• Compare expression across multiple tissue types, considering both healthy and diseased states.

• Combine with tissue-specific markers to identify precise cellular localization.

• Validate IHC findings with qPCR or Western blot analysis of tissue lysates.

Previous research has suggested C1QTNF3 may play a role in skeletal development , making bone and cartilage tissues of particular interest. Additionally, examining expression patterns under different metabolic conditions can provide insights into the regulatory mechanisms controlling C1QTNF3 expression.

What are common issues with C1QTNF3 antibody experiments and how can they be resolved?

Researchers working with C1QTNF3 antibodies may encounter several common challenges:

For Western blot applications specifically, be aware that multiple isoforms of human CTRP3 have been reported , which may explain detection of multiple bands. Confirming the specificity of observed bands through appropriate controls is essential for accurate data interpretation.

How should C1QTNF3 data be normalized and analyzed across different experimental models?

Proper normalization is crucial for meaningful comparison of C1QTNF3 data across different samples and experimental models:

• For Western blot analysis:

  • Use consistent loading controls (β-actin, GAPDH, etc.)

  • Consider tissue-specific housekeeping genes when comparing across tissues

  • Present data as fold-change relative to appropriate controls

• For ELISA quantification:

  • Generate standard curves for each experimental run

  • Consider normalizing to total protein concentration for tissue or cell lysates

  • For serum/plasma samples, ensure consistent collection and processing protocols

• For IHC analysis:

  • Use digital image analysis with consistent thresholding

  • Include negative controls (secondary antibody only) and positive controls

  • Consider semi-quantitative scoring by multiple observers

When comparing data across species, remember that human and mouse CORS26 share approximately 95% amino acid sequence homology , but expression patterns and regulation may differ, requiring careful interpretation of cross-species comparisons.

How can C1QTNF3 antibodies be utilized in emerging research areas?

C1QTNF3 antibodies offer potential for exploring several emerging research areas:

• Metabolic Disease Models:

  • Investigate CTRP3's role in regulating hepatic glucose output in diabetes models

  • Study changes in CTRP3 expression and signaling in obesity and insulin resistance

  • Examine the therapeutic potential of modulating CTRP3 levels

• Skeletal Development Studies:

  • Explore CTRP3's suggested role in bone and cartilage development

  • Study expression patterns during different stages of skeletal formation

  • Investigate potential interactions with other factors involved in skeletogenesis

• Reproductive Endocrinology:

  • Further characterize the role of CTRP3 in testosterone production

  • Investigate potential contributions to reproductive disorders

  • Study gender-specific differences in CTRP3 expression and function

When designing experiments in these areas, combining antibody-based detection methods with functional assays and genetic approaches will provide the most comprehensive understanding of C1QTNF3 biology and potential applications.

What technological advances might improve C1QTNF3 antibody applications?

Emerging technologies and approaches may enhance C1QTNF3 antibody applications:

• Single-cell analysis techniques combined with C1QTNF3 antibodies could reveal cell-type specific expression patterns within heterogeneous tissues.

• Proximity labeling methods (BioID, APEX) using C1QTNF3 antibodies may identify novel interaction partners in different cellular contexts.

• Advanced imaging techniques such as super-resolution microscopy or expansion microscopy could provide detailed insights into subcellular localization of C1QTNF3.

• Development of phospho-specific antibodies for C1QTNF3 and its downstream targets would enhance signaling pathway studies.

• CRISPR-engineered cellular and animal models with tagged endogenous C1QTNF3 could facilitate antibody-based studies with reduced concerns about specificity.

Researchers interested in contributing to these technological advances should consider validating new C1QTNF3 antibody applications against appropriate controls, including knockout/knockdown systems.