TAC1 Antibody, HRP conjugated

What is TAC1 and why is it significant in neuroscience research?

TAC1 (Protachykinin-1) is a precursor protein that is cleaved into multiple bioactive tachykinin peptides, including Substance P, Neurokinin A (NKA), Neuropeptide K (NPK), and Neuropeptide gamma. These peptides function as neurotransmitters that interact with nerve receptors and smooth muscle cells . TAC1-derived peptides are particularly significant in neuroscience research because they excite neurons, evoke behavioral responses, act as potent vasodilators and secretagogues, and contract smooth muscles either directly or indirectly . The gene encoding TAC1 (also sometimes referred to as TAC2 in older literature) is essential for understanding neuropeptide signaling pathways involved in pain perception, neuroinflammation, and various neurological conditions .

What are the key characteristics of TAC1 Antibody, HRP conjugated?

TAC1 Antibody, HRP conjugated is a polyclonal antibody raised in rabbits against recombinant human Protachykinin-1 protein (specifically amino acids 20-129) . The antibody has been directly conjugated with horseradish peroxidase (HRP) enzyme, creating a molecular complex that enables direct detection without requiring secondary antibodies . Key features include:

• Host Species: Rabbit

• Clonality: Polyclonal

• Isotype: IgG

• Target Specificity: Human TAC1

• Applications: Primarily ELISA

• Immunogen: Recombinant Human Protachykinin-1 protein (20-129AA)

• Conjugation: HRP (Horseradish Peroxidase)

• Buffer Composition: 0.03% Proclin 300, 50% Glycerol, 0.01M PBS, pH 7.4

How does HRP conjugation benefit immunodetection techniques?

HRP conjugation provides several methodological advantages for researchers:

• Direct Detection: The HRP enzyme is directly attached to the primary antibody, eliminating the need for secondary antibody incubation steps, which reduces protocol time and potential sources of background noise .

• Enhanced Sensitivity: HRP catalyzes reactions that produce colorimetric, chemiluminescent, or fluorescent signals, enhancing detection sensitivity when combined with appropriate substrates .

• Stable Conjugation: The molecular complex formed between antibody and HRP is stable (approximately 400,000 daltons as estimated by gel chromatography for similar conjugates), providing reliable detection across multiple experimental conditions .

• Versatility: HRP-conjugated antibodies can be used across multiple detection platforms including ELISA, immunohistochemistry, and Western blotting with appropriate optimization .

The conjugation process typically results in antibody:HRP ratios of approximately 1:1 to 1:4, creating a reagent with molecular weights between approximately 190,000-325,000 daltons when considering the antibody (~150 kDa) and HRP (~44 kDa) components .

What are the optimal conditions for using TAC1 Antibody, HRP conjugated in ELISA?

Based on the technical information available for TAC1 Antibody, HRP conjugated, researchers should consider the following optimization parameters for ELISA applications:

Sample Preparation and Assay Conditions:

• Antibody Dilution Range: Start with manufacturer-recommended dilutions (typically 1:100 to 1:1000 for HRP-conjugated antibodies)

• Incubation Temperature: 37°C for standard protocols; 12°C for extended incubations may improve sensitivity based on similar conjugated antibodies

• Incubation Time: 1-2 hours for standard protocols; overnight incubations at lower temperatures may enhance signal

• Buffer System: PBS with 0.05% Tween-20 is typically used for washing steps

• Blocking Agent: 1-5% BSA or non-fat milk in PBS is recommended

Signal Development:

• Substrate Options: TMB (3,3',5,5'-Tetramethylbenzidine) for colorimetric detection

• Detection Time: Monitor signal development for 5-30 minutes depending on substrate

• Signal Stopping: 2N H₂SO₄ or equivalent stopping solution when appropriate color develops

For validation, include appropriate positive and negative controls, and consider a titration series to determine optimal antibody concentration for your specific sample type.

How can I troubleshoot weak or non-specific signals when using TAC1 Antibody, HRP conjugated?

When encountering weak or non-specific signals, consider implementing the following troubleshooting strategies:

For Weak Signals:

• Increase Antibody Concentration: Try using a higher concentration of the TAC1 Antibody, HRP conjugated within the recommended range

• Extend Incubation Time: Longer incubation periods, especially at lower temperatures (4-12°C), may enhance specific binding

• Add Detergent: Including 0.1% Triton X-100 in the incubation buffer can improve tissue penetration and binding efficacy, as demonstrated with similar HRP-conjugated antibodies

• Optimize Antigen Retrieval: For tissue-based applications, ensure proper antigen retrieval methods are employed

• Check Storage Conditions: Ensure the antibody has been stored according to manufacturer recommendations (-20°C or -80°C, avoiding freeze-thaw cycles)

For Non-specific Signals:

• Increase Blocking: Use more stringent blocking conditions (5% BSA or non-fat milk)

• Adjust Washing: Incorporate additional or longer washing steps

• Dilute Antibody: If background is high, increase the dilution factor of the antibody

• Pre-absorb Antibody: Consider pre-absorbing the antibody with the immunizing peptide to validate specificity

• Review Buffer Components: The presence of 50% glycerol in the antibody formulation can affect binding in some applications; dialysis may be considered if necessary

Can TAC1 Antibody, HRP conjugated be used for applications beyond ELISA?

While the TAC1 Antibody, HRP conjugated is primarily validated for ELISA , evidence from similar HRP-conjugated antibodies suggests potential utility in other applications with appropriate optimization:

Immunohistochemistry (IHC):
Researchers have successfully used HRP-conjugated antibodies for direct immunocytochemistry of substance P (a product of TAC1) . Based on this precedent, TAC1 Antibody, HRP conjugated might be adaptable for IHC with the following considerations:

• Fixation: Paraformaldehyde fixation (4%) is typically used

• Tissue Processing: Paraffin-embedded or frozen sections (10-20 μm)

• Permeabilization: Include 0.1% Triton X-100 to improve tissue penetration

• Incubation Conditions: Extended incubations (24-48 hours) at 12°C may yield optimal results

• Signal Development: DAB (3,3'-Diaminobenzidine) substrate system for visualization

Western Blotting:
Adaptation for Western blotting might be possible based on protocols used for other HRP-conjugated antibodies :

• Sample Preparation: Standard protein extraction and denaturation

• Transfer Conditions: PVDF membrane recommended for optimal protein binding

• Blocking: 5% non-fat milk or BSA in TBST

• Antibody Dilution: Start with 1:1000 to 1:5000 dilutions

• Detection: Enhanced chemiluminescence (ECL) substrates for optimal sensitivity

Validation experiments are essential before using TAC1 Antibody, HRP conjugated for applications beyond its validated use in ELISA.

How can TAC1 Antibody, HRP conjugated be employed in studying neuroinflammatory processes?

TAC1-encoded peptides, particularly Substance P, play significant roles in neuroinflammation. Researchers investigating these processes can utilize TAC1 Antibody, HRP conjugated with the following experimental approaches:

Neuroinflammation Models:

• Cell Culture Systems: Detect TAC1 expression in neuronal-glial co-cultures following inflammatory stimuli (LPS, TNF-α, IL-1β)

• Tissue Analysis: Quantify TAC1 peptide levels in brain tissue from neuroinflammatory disease models

• Cellular Localization: Determine TAC1 distribution in activated microglia, astrocytes, and neurons during inflammatory responses

Methodological Considerations:

• Sample Preparation: For brain tissue, careful extraction protocols should be employed to preserve tachykinin peptides, which can be degraded by endogenous proteases

• Detection Strategy: Sandwich ELISA using TAC1 Antibody, HRP conjugated as the detection antibody paired with a capture antibody against a different TAC1 epitope

• Quantification: Standard curves using recombinant TAC1 peptides (Substance P, Neurokinin A) for accurate quantification

• Controls: Include peptide-specific blocking controls to confirm specificity of detection in complex tissue samples

What strategies can be used to study the relationship between TAC1 expression and cancer progression?

TAC1 and its peptide products have been implicated in cancer biology, particularly in aspects of tumor growth, angiogenesis, and metastasis. Researchers investigating these connections can implement the following approaches:

Experimental Designs:

• Expression Analysis: Compare TAC1 levels between tumor tissues and adjacent normal tissues using ELISA with TAC1 Antibody, HRP conjugated

• Cell Line Profiling: Screen cancer cell lines for differential TAC1 expression under various conditions (hypoxia, drug treatment)

• Functional Studies: Correlate TAC1 expression with cancer cell proliferation, migration, and invasion capabilities

Research Applications Table:

When designing these studies, researchers should consider that TAC1 gene products include multiple peptides (Substance P, Neurokinin A, NPK, Neuropeptide gamma) , and the TAC1 Antibody, HRP conjugated may detect the precursor protein and/or processed peptides depending on the epitope recognized.

How can computational approaches complement experimental studies using TAC1 Antibody, HRP conjugated?

Integrating computational methods with experimental data generated using TAC1 Antibody, HRP conjugated can significantly enhance research outcomes:

Bioinformatic Strategies:

• Sequence Analysis: Examine the epitope recognized by the antibody (within amino acids 20-129 of human TAC1) for evolutionary conservation across species

• Structural Modeling: Predict the three-dimensional structure of TAC1 peptides and identify surface-exposed regions accessible to antibody binding

• Network Analysis: Map TAC1 interactions with receptors (NK1R, NK2R, NK3R) and downstream signaling pathways

Data Integration Approaches:

• Multi-omics Integration: Correlate TAC1 protein levels (detected via TAC1 Antibody, HRP conjugated) with transcriptomic and metabolomic data

• Pathway Enrichment: Identify biological processes and signaling cascades associated with differential TAC1 expression

• Machine Learning: Develop predictive models incorporating TAC1 expression data for disease progression or treatment response

Validation Strategy for Computational Predictions:

• Generate hypotheses based on computational models

• Test predictions using TAC1 Antibody, HRP conjugated in relevant experimental systems

• Refine models based on experimental feedback

• Develop more targeted research questions based on integrated insights

What are the key storage and handling parameters to maintain optimal activity of TAC1 Antibody, HRP conjugated?

Proper storage and handling are critical for maintaining the activity and specificity of TAC1 Antibody, HRP conjugated:

Storage Recommendations:

• Temperature: Store at -20°C or -80°C for long-term preservation

• Aliquoting: Upon receipt, prepare small working aliquots to avoid repeated freeze-thaw cycles

• Thawing: Thaw aliquots gradually on ice before use

• Short-term Storage: For ongoing experiments, store at 2-8°C for up to one week

• Stability: HRP conjugates typically remain stable for 6-12 months when stored properly

Handling Guidelines:

• Buffer Compatibility: The antibody is formulated in a buffer containing 50% glycerol, 0.01M PBS (pH 7.4), and 0.03% Proclin 300 as a preservative

• Dilution Media: When diluting, use buffers compatible with the original formulation (PBS-based)

• Temperature Sensitivity: Avoid exposure to temperatures above 25°C, which can denature both the antibody and the HRP enzyme

• Light Sensitivity: Minimize exposure to direct light, as HRP conjugates can be photosensitive

• Contamination Prevention: Use sterile technique when handling to prevent microbial contamination

How can I validate the specificity of TAC1 Antibody, HRP conjugated in my experimental system?

Validating antibody specificity is crucial for generating reliable research data. For TAC1 Antibody, HRP conjugated, consider these validation approaches:

Positive Controls:

• Recombinant Protein: Use purified recombinant human TAC1 protein as a positive control

• Known Positive Tissues/Cells: Test tissues with documented TAC1 expression (e.g., submucosal plexus in human colon)

• Overexpression Systems: Analyze cells transfected to overexpress TAC1

Negative Controls:

• Peptide Blocking: Pre-incubate antibody with excess immunizing peptide to confirm signal specificity

• Genetic Models: Test samples from TAC1 knockout models (if available)

• Low-expressing Tissues: Examine tissues known to express minimal TAC1

Specificity Validation Table:

What are the considerations for developing multiplex assays that include TAC1 Antibody, HRP conjugated?

Multiplexing allows simultaneous detection of multiple targets, enhancing experimental efficiency and enabling correlation analyses. When incorporating TAC1 Antibody, HRP conjugated into multiplex assays, consider:

Assay Design Principles:

• Enzyme Selection: Since this antibody is HRP-conjugated, other targets would need different enzyme conjugates (e.g., alkaline phosphatase) or fluorescent labels

• Substrate Compatibility: Choose HRP substrates with spectral properties distinct from other detection systems

• Cross-reactivity Testing: Verify that other antibodies in the multiplex panel don't cross-react with TAC1 or the TAC1 Antibody

Multiplex Configurations:

• Spatial Separation: Immobilize different capture antibodies in distinct locations (e.g., different wells or array spots)

• Spectral Separation: Use HRP substrates with different emission spectra from other detection systems

• Temporal Separation: Apply detection reagents sequentially with intermediate blocking steps

Experimental Considerations:

• Signal Balance: Adjust antibody concentrations to achieve comparable signal intensities across targets

• Internal Controls: Include standardization controls for each detection system

• Data Analysis: Apply appropriate normalization methods when comparing signals from different detection systems

• Validation: Confirm that multiplex results match those obtained from single-plex assays

How can TAC1 Antibody, HRP conjugated contribute to pain research?

Substance P and other TAC1-derived peptides are critical mediators in pain signaling pathways. TAC1 Antibody, HRP conjugated can be applied in pain research contexts:

Experimental Models:

• Inflammatory Pain: Quantify TAC1 peptide expression in dorsal root ganglia and spinal cord following inflammatory stimuli

• Neuropathic Pain: Compare TAC1 levels before and after nerve injury

• Chronic Pain Conditions: Assess TAC1 expression changes in response to analgesic treatments

Methodological Approach:

• Tissue Preparation: Process neural tissues (dorsal root ganglia, spinal cord, brain) with careful attention to preserving peptide integrity

• Quantification: Develop standard curves using recombinant TAC1 peptides to accurately measure concentrations

• Comparative Analysis: Correlate TAC1 expression with behavioral pain assessments and other molecular markers

What role can TAC1 detection play in understanding gastrointestinal disorders?

TAC1-derived peptides, particularly Substance P, are important in regulating gastrointestinal motility, secretion, and inflammation. TAC1 Antibody, HRP conjugated can be utilized to investigate GI disorders:

Research Applications:

• Inflammatory Bowel Disease: Compare TAC1 expression between healthy and inflamed intestinal tissues

• Irritable Bowel Syndrome: Assess TAC1 levels in patient biopsies and correlate with symptom severity

• Enteric Nervous System Function: Examine TAC1 distribution in submucosal and myenteric plexuses

Sample Types and Processing:

• Tissue Biopsies: Process fresh or frozen GI tissue samples for TAC1 quantification

• Intestinal Organoids: Analyze TAC1 expression in 3D culture models of intestinal epithelium

• Enteric Neurons: Isolate and culture enteric neurons to study TAC1 regulation

How might TAC1 Antibody, HRP conjugated be used in respiratory research?

TAC1-derived peptides influence airway function through neurogenic inflammation, bronchoconstriction, and mucus secretion. TAC1 Antibody, HRP conjugated can support respiratory research:

Research Directions:

• Asthma Models: Quantify TAC1 in bronchial tissues following allergen challenge

• COPD Studies: Compare TAC1 expression between smokers and non-smokers

• Neurogenic Inflammation: Track TAC1 changes during respiratory inflammatory processes

Experimental Design Table:

By addressing these frequently asked questions, researchers can optimize their use of TAC1 Antibody, HRP conjugated across various experimental contexts, from basic characterization to advanced disease-specific applications.