C1QTNF9 Antibody, Biotin conjugated

What is C1QTNF9 and how is it structurally organized?

C1QTNF9 belongs to the highly conserved CTRP family of secreted proteins, which currently includes 15 identified members (CTRP1-15). The protein features a modular organization with four distinct domains: an N-terminal signal peptide, a domain with conserved cysteine residues, a collagen-like domain containing Gly-X-Y repeats, and a C-terminal globular C1q domain. This 40 kDa glycoprotein contains multiple hydroxylated proline residues in its collagenous region and circulates as homotrimers, higher-order multimers, and heterotrimers with adiponectin .

While all CTRPs are structurally related to adiponectin (an insulin-sensitizing hormone secreted from adipose tissue), C1QTNF9 has its own unique expression profile and non-redundant functions. It is preferentially expressed in adipose tissue and plays important roles in glucose homeostasis .

What is the difference between C1QTNF9 and CTRP9?

These are actually the same protein. C1QTNF9 is the official gene/protein name (C1q and Tumor Necrosis Factor Related Protein 9), while CTRP9 is a commonly used alternative designation. Other synonyms include C1QTNF9A and AQL1. In the literature, you'll find both designations used interchangeably, but they refer to the same biological molecule .

What model organisms are appropriate for C1QTNF9 research?

Human and mouse models are the most commonly studied for C1QTNF9 research. The human C1QTNF9 shares approximately 85% amino acid sequence identity with mouse and rat C1QTNF9, making these species suitable for comparative studies . Commercially available antibodies and detection kits are specifically designed for either human or mouse C1QTNF9, so researchers should select the appropriate species-specific reagents for their experimental models .

What is the principle behind biotin-conjugated antibody detection of C1QTNF9?

Biotin-conjugated antibodies are essential components of the sandwich ELISA method used to detect C1QTNF9. In this assay, the microtiter plate is pre-coated with an antibody specific to C1QTNF9. When standards or samples are added to the wells, C1QTNF9 binds to this immobilized antibody. Subsequently, a biotin-conjugated antibody specific to C1QTNF9 is added, which binds to the captured C1QTNF9. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added, which binds to the biotin. After TMB substrate addition, only wells containing C1QTNF9, biotin-conjugated antibody, and enzyme-conjugated Avidin exhibit a color change. The reaction is terminated with sulfuric acid solution, and color intensity is measured spectrophotometrically at 450nm (±10nm). The concentration of C1QTNF9 is determined by comparing sample OD values to a standard curve .

What sample types can be effectively analyzed using biotin-conjugated C1QTNF9 antibodies?

Biotin-conjugated C1QTNF9 antibodies can be used to analyze multiple biological sample types including:

For optimal results, proper sample preparation is essential to minimize interference from other components in the biological matrices .

What are the critical parameters for optimal C1QTNF9 detection using biotin-conjugated antibodies?

When working with biotin-conjugated antibodies for C1QTNF9 detection, researchers should carefully control the following parameters:

• Antibody concentration: The biotin-conjugated antibody must be diluted to the optimal working concentration using the appropriate buffer.

• Incubation conditions: Temperature (typically 37°C) and time must be strictly controlled.

• Washing steps: Thorough washing between steps is crucial to reduce background.

• Detection sensitivity: The minimum detection limit for human C1QTNF9 is around 0.469-0.781 ng/mL, while for mouse C1QTNF9 it's approximately 6.3 pg/mL.

• Detection range: For human assays, the typical range is 0.781-50 ng/mL; for mouse assays, it's 15.63-1000 pg/mL.

Precision parameters also matter, with intra-assay precision (CV%) typically <8% and inter-assay precision (CV%) <10% .

How can C1QTNF9 be studied in the context of diabetic cardiomyopathy?

Recent research has investigated C1QTNF9's role in diabetic cardiomyopathy using adeno-associated virus (AAV) vector systems. In these studies, the C1qtnf9 gene was cloned into AAV-vector genome plasmids under the control of tissue-specific promoters (like TnT for cardiac-specific expression). The vectors are then administered to experimental animals (typically mice) via intravenous injection with dosages around 1 × 10^12 viral genomes. Control groups receive AAV vectors expressing reporter genes like renilla luciferase (rluC) under the same promoter.

Following vector administration, animals are typically maintained on experimental diets for 12 weeks before being sacrificed for analysis. This approach allows researchers to study the protective effects of C1QTNF9 overexpression against diabetic cardiac damage .

What are the key considerations for interpreting C1QTNF9 cross-reactivity data?

When evaluating antibody specificity, researchers should be aware that despite high sensitivity and specificity of commercial C1QTNF9 detection assays, cross-reactivity with analogues can occur. Current limitations in knowledge make it difficult to comprehensively assess cross-reactivity between C1QTNF9 and all its analogues.

For rigorous experimental design, researchers should:

• Include proper controls to account for potential cross-reactivity

• Validate findings using complementary methods

• Consider the structural similarity between C1QTNF9 and other CTRP family members

• Be particularly cautious when studying tissues with high expression of multiple CTRP family members

Documented cross-reactivity data should be consulted for the specific antibody being used, and researchers should acknowledge this limitation in their experimental interpretation .

How does C1QTNF9 interact with adiponectin and what implications does this have for experimental design?

C1QTNF9 not only forms homotrimers and higher-order multimers but also forms heterotrimers with adiponectin. This interaction has significant implications for experimental design:

• When studying C1QTNF9 function, researchers must consider its interaction with adiponectin

• Immunoprecipitation experiments targeting C1QTNF9 may co-precipitate adiponectin

• Functional studies should distinguish between effects mediated by C1QTNF9 homotrimers versus C1QTNF9-adiponectin heterotrimers

• Expression analyses should consider the co-regulation of adiponectin and C1QTNF9

These protein-protein interactions affect downstream signaling pathways and should be carefully considered when interpreting experimental results related to glucose homeostasis and metabolic regulation .

What are the most common issues encountered with biotin-conjugated C1QTNF9 antibodies and how can they be resolved?

Researchers should also maintain consistent sample preparation methods and ensure all reagents are at room temperature before use .

What storage and handling precautions are necessary for maintaining biotin-conjugated antibody activity?

To maintain optimal antibody performance:

• Store reconstituted antibodies at -20 to -70°C for long-term storage (up to 6 months)

• For short-term use (up to 1 month), store at 2-8°C under sterile conditions

• Avoid repeated freeze-thaw cycles by aliquoting antibodies before freezing

• Use manual defrost freezers rather than auto-defrost ones

• Prepare working dilutions freshly before use

• Protect biotin-conjugated antibodies from prolonged exposure to light

Following these guidelines will help maintain antibody integrity and experimental reproducibility .

How should standard curves be generated and validated for C1QTNF9 quantification?

For accurate C1QTNF9 quantification:

• Prepare a series of standards through serial dilution (e.g., 1000, 500, 250, 125, 62.5, 31.25, 15.63 pg/mL for mouse C1QTNF9)

• Plot the standard curve using OD values against known concentrations

• Apply appropriate curve-fitting methods (four-parameter logistic curve fit is recommended)

• Validate the standard curve by calculating recovery of standards (acceptable range: 80-120%)

• Assess linearity by testing dilutions of high-concentration samples

• Include quality control samples of known concentration on each plate

The coefficient of determination (R²) should exceed 0.99 for a reliable standard curve. Sample concentrations should be calculated by interpolation from the standard curve, accounting for any dilution factors .

What statistical approaches are appropriate for analyzing C1QTNF9 expression data across different experimental conditions?

When analyzing C1QTNF9 expression data:

• For comparing two groups: Use paired or unpaired t-tests (depending on study design)

• For multiple group comparisons: Apply one-way ANOVA followed by appropriate post-hoc tests (Tukey's, Bonferroni, etc.)

• For time-course studies: Consider repeated measures ANOVA or mixed models

• For correlation with other parameters: Calculate Pearson's or Spearman's correlation coefficients

Data normality should be verified before parametric testing. For non-normally distributed data, non-parametric alternatives should be employed. Additionally, researchers should account for covariates that might influence C1QTNF9 expression (age, sex, BMI) through multiple regression or ANCOVA when appropriate. Sample size calculations should be performed during experimental planning to ensure adequate statistical power .

What emerging technologies could enhance C1QTNF9 detection and functional analysis?

Emerging technologies for C1QTNF9 research include:

• Single-cell RNA sequencing to identify cell-specific expression patterns

• CRISPR-Cas9 gene editing for creating precise C1QTNF9 modifications

• Proximity ligation assays to study C1QTNF9 protein-protein interactions in situ

• Advanced imaging techniques including super-resolution microscopy for subcellular localization

• Proteomics approaches to identify post-translational modifications

• Computational modeling to predict C1QTNF9 structure-function relationships

These technologies may provide deeper insights into C1QTNF9 biology beyond what traditional antibody-based detection methods can offer .

How might C1QTNF9 research contribute to therapeutic developments for metabolic and cardiovascular diseases?

Given C1QTNF9's role in glucose homeostasis and cardioprotection, several therapeutic avenues could emerge:

• Development of recombinant C1QTNF9 as a potential therapeutic for metabolic disorders

• Creation of synthetic peptides mimicking C1QTNF9's active domains

• Identification of small molecules that enhance endogenous C1QTNF9 expression or activity

• Gene therapy approaches using AAV vectors for tissue-specific C1QTNF9 expression

• Combinatorial approaches targeting C1QTNF9 and adiponectin simultaneously

Research using biotin-conjugated antibodies will remain crucial for assessing C1QTNF9 levels in preclinical and clinical studies evaluating these potential therapeutic strategies .