TAC1 Antibody, HRP conjugated

What is TAC1 and why is it important in research?

TAC1, also known as Protachykinin-1, is a gene that encodes for a family of neuropeptides known as tachykinins. These peptides are characterized by their ability to rapidly stimulate contraction of intestinal muscle, hence the name "tachykinins" . TAC1 is responsible for four products of the tachykinin peptide hormone family: substance P, neurokinin A (NKA), neuropeptide K (NPK), and neuropeptide gamma . These peptides function as active molecules that excite neurons, evoke behavioral responses, act as potent vasodilators and secretagogues, and contract many smooth muscles (directly or indirectly) . TAC1 and its peptides are widely distributed in the nervous system and play important roles in neurotransmission, neuromodulation, and neuroinflammation, making them significant targets in neuroscience, cancer, and metabolism research .

What is the molecular structure and characteristics of TAC1?

TAC1 protein has a calculated molecular weight of 15 kDa (129 amino acids), though it is sometimes observed at 17 kDa in experimental conditions . The human TAC1 gene has the GenBank Accession Number BC018047 and NCBI Gene ID 6863 . The protein's UniProt ID is P20366 . The gene is evolutionarily conserved and ubiquitously expressed, although its functions are organ-specific . Structurally, TAC1 functions as a precursor protein that is cleaved to produce the bioactive tachykinin peptides, with the full sequence spanning 129 amino acids in humans .

What are the key differences between TAC1 antibodies with different conjugates?

The primary difference lies in their detection methods and applications:

• HRP-conjugated TAC1 antibodies: These antibodies have horseradish peroxidase directly linked to them, enabling colorimetric detection in assays like ELISA without requiring a secondary antibody . The enzymatic activity of HRP produces a color change when exposed to an appropriate substrate.

• Unconjugated TAC1 antibodies: These require a labeled secondary antibody for detection and are versatile for applications like IHC, IF/ICC, and Western blotting .

• FITC-conjugated TAC1 antibodies: These are directly labeled with a fluorescent tag for fluorescence-based detection methods .

• Biotin-conjugated TAC1 antibodies: These utilize biotin-avidin interactions for signal amplification and are particularly useful in sensitive detection methods .

How does the expression pattern of TAC1 differ across tissues?

TAC1 expression shows distinct tissue distribution patterns. α-TAC1 mRNA is localized in numerous neurons of the brain, while β-TAC1 and γ-TAC1 are predominantly expressed in intrinsic enteric neurons and sensory neurons . According to immunohistochemistry validation data, TAC1 protein is positively detected in mouse brain tissue, human brain tissue, human hypothalamus tissue, human stomach tissue, and human pancreas tissue . Additionally, TAC1 has been detected in PC-12 cells using immunofluorescence techniques . Understanding this distribution pattern is crucial for designing appropriate controls when working with TAC1 antibodies.

What are the validated applications for TAC1 Antibody, HRP conjugated?

TAC1 Antibody, HRP conjugated is primarily validated for ELISA applications . The antibody shows reactivity with human samples and has been specifically optimized for competitive inhibition ELISA reactions . Some TAC1 antibody formulations, though not necessarily all HRP-conjugated versions, are also validated for other applications including:

It's important to note that while HRP-conjugated versions are optimized for ELISA, the appropriate application should be verified for each specific antibody formulation .

How do I optimize a competitive inhibition ELISA using TAC1 Antibody, HRP conjugated?

For a competitive inhibition ELISA using TAC1 Antibody, HRP conjugated:

• Assay Principle: A competitive inhibition reaction is launched between biotin-labeled TAC1 and unlabeled TAC1 (standards or samples) with the pre-coated antibody specific to TAC1 .

• Protocol Optimization:

  • Coating Concentration: Use optimal concentration of capture antibody (typically 1-10 μg/ml)

  • Sample Preparation: Ensure proper dilution of samples in appropriate buffer

  • Incubation Time and Temperature: Standard conditions are 37°C for 1-2 hours

  • Blocking: Use 1-5% BSA in PBS to reduce background

  • Washing: Thorough washing between steps is critical (3-5 washes)

  • Substrate Addition: Add TMB substrate and monitor color development

  • Stop Solution: Add stop solution when appropriate color intensity is reached

• Data Analysis: Generate a standard curve using known concentrations of TAC1. The intensity of color developed is inversely proportional to the concentration of TAC1 in the sample .

• Optimization Tips:

  • Always run parallel positive and negative controls

  • Titrate the antibody in each testing system to obtain optimal results (starting with manufacturer's recommended dilution)

  • Sample-dependent variations may require additional optimization

What are the critical buffer components for maintaining TAC1 Antibody, HRP conjugated activity?

The optimal storage buffer composition for maintaining TAC1 Antibody, HRP conjugated activity typically includes:

• Base Buffer: PBS at pH 7.3-7.4 as the primary buffer component

• Preservative: 0.02-0.03% Proclin 300 or sodium azide to prevent microbial growth

• Stabilizer: 50% Glycerol to maintain protein stability during freeze-thaw cycles

• Additional Components: Some formulations contain 0.1% BSA for added stability

When designing experimental buffers for working with TAC1 antibodies, consider:

• Avoiding detergents that could interfere with antigen-antibody interactions

• Maintaining pH between 7.2-7.4 for optimal binding

• Using freshly prepared buffers to ensure consistency in results

• Avoiding repeated freeze-thaw cycles that could degrade the antibody

How can TAC1 Antibody, HRP conjugated be used to study SDF-1α-mediated regulation of TAC1 expression?

Based on research findings, SDF-1α (Stromal cell-derived growth factor-1α) has been shown to regulate TAC1 expression in bone marrow stroma through a concentration-dependent mechanism . To investigate this relationship:

• Experimental Design:

  • Treat bone marrow stromal cells with varying concentrations of SDF-1α (e.g., 20, 50, and 100 ng/ml)

  • After appropriate incubation periods, conduct competitive ELISA using TAC1 Antibody, HRP conjugated to quantify TAC1 peptide production (particularly substance P)

  • Compare with reporter gene assays and Northern blot analyses to correlate protein levels with transcriptional activity and mRNA expression

• Expected Results:

  • A bell-shaped response curve where 20 ng/ml SDF-1α stimulates TAC1 expression

  • Higher concentrations (50 and 100 ng/ml) inhibit TAC1 expression

  • NF-κB involvement in the repressive effects at higher concentrations of SDF-1α

• Significance:
  This approach helps elucidate the neural-immune-hemopoietic axis, particularly how SDF-1α levels above baseline in bone marrow stroma induce substance P production to stimulate hemopoiesis, while substance P does not act as an autocrine stimulator of SDF-1α production .

What techniques can be combined with TAC1 antibody detection to study its role in neuroinflammation?

To comprehensively study TAC1's role in neuroinflammation, researchers can employ multiple complementary approaches:

• Multiplex Immunoassays:

  • Combine TAC1 Antibody, HRP conjugated ELISA with multiplexed cytokine/chemokine assays

  • Correlate TAC1 peptide levels with inflammatory mediators in the same samples

• Tissue-Specific Analysis:

  • Use unconjugated TAC1 antibodies for IHC/IF on brain tissue sections (dilution 1:50-1:500)

  • Co-stain with markers for inflammatory cells (microglia, astrocytes)

  • Perform spatial analysis of TAC1 expression relative to inflammatory foci

• Cell-Specific Investigation:

  • Employ flow cytometry with appropriate TAC1 antibodies to identify TAC1-expressing cell populations

  • Sort cells for RNA-seq to identify associated inflammatory gene networks

• Functional Assessments:

  • Combine TAC1 quantification with neurobehavioral testing in animal models

  • Correlate substance P levels (detected using TAC1 Antibody, HRP conjugated) with markers of blood-brain barrier integrity

• Receptor Interaction Studies:

  • Investigate TAC1 peptide interactions with their receptors (e.g., neurokinin 1 rather than neurokinin 2)

  • Link receptor activation to downstream inflammatory signaling pathways

How can I design experiments to investigate the relationship between TAC1 and hemopoiesis?

To investigate the relationship between TAC1 and hemopoiesis:

• Long-term Culture-Initiating Cell Assays:

  • Establish bone marrow stromal cell cultures

  • Manipulate TAC1 expression (knockdown/overexpression)

  • Quantify TAC1 peptide production using TAC1 Antibody, HRP conjugated

  • Assess hemopoietic supporting capacity of stromal cells under different conditions

• SDF-1α-Mediated Effects:

  • Treat cultures with varying concentrations of SDF-1α (20, 50, 100 ng/ml)

  • Measure TAC1 peptide production (particularly substance P)

  • Correlate with hemopoietic activity

• Receptor Studies:

  • Use specific receptor antagonists (e.g., for neurokinin 1 receptor)

  • Determine which receptor mediates the hemopoietic effects of TAC1 peptides

• Intracellular Signaling Analysis:

  • Investigate NF-κB pathway involvement in TAC1 regulation

  • Use reporter gene assays with the 5' flanking region of TAC1

  • Correlate with Northern analyses and ELISA for mRNA and protein levels

This approach would build on findings that SDF-1α affects hemopoiesis indirectly through substance P production, with neurokinin 1 as the relevant receptor, while substance P does not regulate SDF-1α production in stroma .

What are common issues when using TAC1 Antibody, HRP conjugated in ELISA, and how can they be resolved?

Common issues and their solutions include:

For optimal results:

• Store antibody at -20°C or -80°C and avoid repeated freeze-thaw cycles

• Prepare working dilutions immediately before use

• Include appropriate positive controls (e.g., mouse brain tissue, human brain/hypothalamus tissue)

• For antigen retrieval, use TE buffer pH 9.0 or citrate buffer pH 6.0 as recommended

How do I interpret contradictory results between TAC1 protein levels and gene expression data?

When facing contradictions between TAC1 protein levels (measured by ELISA with TAC1 Antibody, HRP conjugated) and gene expression data:

• Consider Post-transcriptional Regulation:

  • TAC1 undergoes alternative splicing to produce different isoforms (α, β, γ, δ)

  • Verify which isoform your antibody detects (some may recognize precursors but not processed peptides)

  • Examine microRNA-mediated regulation that might affect translation efficiency

• Protein Processing and Stability:

  • TAC1 is a precursor protein cleaved into multiple bioactive peptides

  • Different antibodies may recognize different epitopes/processing states

  • Assess protease activity in your experimental system

• Temporal Considerations:

  • Gene expression changes often precede protein level changes

  • Design time-course experiments to capture this relationship

  • The SDF-1α study showed correlations among reporter gene activities, mRNA levels (β-preprotachykinin I), and protein levels (substance P)

• Tissue/Cell-Type Specificity:

  • TAC1 expression varies by tissue (brain neurons vs. enteric/sensory neurons)

  • Ensure sampling from the same cell populations for both analyses

  • Consider cell-specific post-translational modifications

• Technical Validation:

  • Cross-validate with multiple antibodies targeting different epitopes

  • Employ both mRNA quantification methods (qPCR, Northern blot) and protein detection methods (ELISA, Western blot)

  • Include positive control tissues with known TAC1 expression (mouse brain, human hypothalamus)

What control samples should I include when working with TAC1 Antibody, HRP conjugated?

To ensure experimental validity when working with TAC1 Antibody, HRP conjugated:

• Positive Controls:

  • Tissue Samples: Mouse brain tissue, human brain/hypothalamus tissue, human stomach tissue, human pancreas tissue (all validated for TAC1 expression)

  • Cell Lines: PC-12 cells (validated for positive IF/ICC detection)

  • Recombinant Protein: Purified TAC1 protein or synthetic TAC1 peptides at known concentrations

• Negative Controls:

  • Antibody Controls: Isotype-matched irrelevant antibody (rabbit IgG)

  • Peptide Neutralization: Pre-absorption of antibody with immunizing peptide

  • Secondary-only Controls: Omit primary antibody to assess non-specific binding

• Technical Controls:

  • Standard Curve: Serial dilutions of recombinant TAC1 protein

  • Dilution Linearity: Serial dilutions of positive samples to verify response proportionality

  • Spike-in Recovery: Addition of known amounts of analyte to verify detection efficiency

• Biological Validation:

  • TAC1 Knockout/Knockdown: Samples from TAC1-deficient models

  • Stimulated Expression: Samples with TAC1 upregulation (e.g., SDF-1α treatment at 20 ng/ml)

  • Cross-species Validation: Verify antibody performance across human, mouse, and rat samples where applicable

How is TAC1 research contributing to understanding the neural-immune-hemopoietic axis?

Research utilizing TAC1 antibodies has revealed crucial insights into the neural-immune-hemopoietic axis:

• SDF-1α as a New Mediator:

  • Studies have demonstrated that SDF-1α regulates TAC1 expression in bone marrow stroma in a concentration-dependent manner

  • At 20 ng/ml, SDF-1α stimulates TAC1 expression, while at 50-100 ng/ml, it inhibits expression through an NF-κB-dependent mechanism

  • This regulation has functional consequences for hemopoiesis, with substance P (a TAC1 peptide) mediating indirect effects of SDF-1α

• Receptor Specificity:

  • Research has identified neurokinin 1, not neurokinin 2, as the relevant receptor mediating TAC1 peptide effects in hemopoiesis

  • This specificity is important for targeted therapeutic interventions

• Unidirectional Regulation:

  • A significant finding is that while SDF-1α regulates substance P production, substance P does not regulate SDF-1α production in stroma

  • This unidirectional relationship clarifies the hierarchy in neural-immune-hemopoietic signaling

These discoveries position TAC1 as a critical link between neural signals, immune responses, and hemopoietic regulation, opening avenues for therapeutic interventions in conditions affecting these systems.

What are the emerging applications of TAC1 antibodies in cancer research?

TAC1 antibodies are increasingly being utilized in cancer research, with several emerging applications:

• Biomarker Development:

  • TAC1 peptides, particularly substance P, are being investigated as potential biomarkers for certain cancers

  • Quantification using TAC1 Antibody, HRP conjugated in ELISA can help establish reference ranges for clinical applications

• Tumor Microenvironment Analysis:

  • TAC1 peptides function in neurogenic inflammation and modulation of immune responses

  • Immunohistochemistry with TAC1 antibodies helps map peptide distribution in tumor microenvironments

  • Co-staining with markers for immune cells, blood vessels, and stromal components provides insights into TAC1's role in tumor biology

• Therapeutic Target Validation:

  • As neurokinin receptor antagonists advance in clinical development, TAC1 antibodies are crucial for validating target expression

  • Correlating TAC1 peptide levels with treatment responses can identify potential predictive biomarkers

• Neuroendocrine Tumor Characterization:

  • TAC1 peptides are expressed in various neuroendocrine tumors

  • Antibody-based detection methods help classify these tumors and understand their biology

These applications leverage TAC1's known roles in neuroinflammation, immune modulation, and vascular responses, which are all relevant to cancer development and progression .

What technical advances are expected to improve TAC1 antibody applications in the near future?

Several technical advances are poised to enhance TAC1 antibody applications:

• Multiplexed Detection Systems:

  • Development of multiplexed platforms allowing simultaneous detection of multiple TAC1 peptides (substance P, neurokinin A, neuropeptide K, neuropeptide gamma)

  • Integration with cytokine/chemokine panels for comprehensive inflammatory profiling

• Improved Specificity:

  • Next-generation antibodies with enhanced specificity for individual TAC1 isoforms and processed peptides

  • Recombinant antibody technology producing more consistent lot-to-lot performance

• Single-Cell Applications:

  • Adaptation of TAC1 antibodies for single-cell proteomics

  • Integration with spatial transcriptomics to correlate protein localization with gene expression at single-cell resolution

• In vivo Imaging:

  • Development of TAC1 antibody derivatives suitable for in vivo imaging

  • Fluorescence or radiotracer-conjugated variants for tracking TAC1 peptide distribution in real-time

• Automated Analysis Platforms:

  • AI-based image analysis tools for quantitative assessment of TAC1 immunostaining patterns

  • Machine learning algorithms to correlate TAC1 expression with clinical outcomes

These advances will enhance the utility of TAC1 antibodies across research applications, from basic neuroscience to clinical oncology and immunology.

What are the detailed specifications of commercially available TAC1 Antibody, HRP conjugated?

Commercially available TAC1 Antibody, HRP conjugated products typically have the following specifications:

How do different antigen retrieval methods affect TAC1 antibody performance in tissue samples?

Antigen retrieval methods can significantly impact TAC1 antibody performance in tissue samples:

• Recommended Methods:

  • Primary recommendation: TE buffer pH 9.0

  • Alternative method: Citrate buffer pH 6.0

• Tissue-Specific Considerations:

  • For brain tissue (mouse and human): TE buffer pH 9.0 provides optimal results

  • For peripheral tissues (human stomach, pancreas): Both methods work, but may require optimization

• Protocol Factors:

  • Temperature: Heat-induced epitope retrieval (95-100°C) typically yields better results than enzymatic methods

  • Duration: 15-20 minutes of heat treatment balances epitope exposure with tissue preservation

  • Cooling: Slow cooling to room temperature improves staining consistency

• Performance Comparison:

  Retrieval Method	Advantages	Limitations	Best For
  TE buffer pH 9.0	Enhanced signal intensity, reduced background, better for membrane epitopes	May cause tissue detachment if not properly optimized	Brain tissue, hypothalamus samples
  Citrate buffer pH 6.0	Gentler on tissue morphology, well-established protocol	Sometimes yields lower signal intensity	Stomach tissue, pancreas tissue, samples with fragile morphology

• Optimization Strategy:

  • Test both methods in parallel with positive control tissues

  • Assess signal-to-noise ratio, staining intensity, and specific localization patterns

  • Titrate antibody dilution (1:50-1:500) for each retrieval method separately

What quality control measures ensure reliable results with TAC1 Antibody, HRP conjugated?

To ensure reliable results with TAC1 Antibody, HRP conjugated, implement the following quality control measures:

• Antibody Validation:

  • Confirm reactivity with positive controls (mouse brain tissue, human brain/hypothalamus tissue)

  • Verify absence of signal in negative controls or with blocking peptide

  • Check lot-to-lot consistency with reference standards

• Assay Performance Metrics:

  • Determine assay sensitivity (limit of detection)

  • Establish standard curve linearity and range

  • Calculate intra-assay and inter-assay coefficients of variation (<10% is desirable)

  • Perform spike-and-recovery tests to assess matrix effects

• Protocol Standardization:

  • Maintain consistent incubation times and temperatures

  • Use calibrated pipettes and validated reagent lots

  • Implement detailed SOPs for each application

• Sample Handling:

  • Standardize collection, processing, and storage conditions

  • Document freeze-thaw cycles for each sample

  • Include stability controls to monitor degradation

• Data Analysis:

  • Use appropriate curve-fitting models for quantitative analysis

  • Include quality control samples in each run (low, medium, high concentrations)

  • Establish acceptance criteria for results validity (e.g., R² > 0.98 for standard curves)

• Documentation:

  • Maintain detailed records of antibody lot, dilution, and storage conditions

  • Document any deviations from standard protocols

  • Archive raw data along with processed results for traceability