NTSR1 Antibody, FITC conjugated

What is NTSR1 and what are its key molecular characteristics?

NTSR1 (Neurotensin Receptor Type 1) is a G-protein coupled receptor that functions as a high-affinity receptor for neurotensin. It has a molecular weight of approximately 43-49 kDa, with search results indicating 43 kDa in Western blot applications and 49 kDa in product specifications . The receptor belongs to the G-protein coupled receptor 1 family and is also known as NT-R-1, NTR1, or NTRH . The receptor's full amino acid sequence begins with MRLNSSAPGTPGTPAADPFQRAQAGLEEALLAPGFGNASGNASERVLAAPSSELDVNTDIYSKVLVTAVY .

When studying NTSR1 using antibodies, researchers should be aware that the receptor's molecular characteristics influence detection methods:

What validated applications exist for NTSR1 antibodies and how might FITC conjugation affect these methods?

Based on search results, NTSR1 antibodies have been validated for several research applications:

FITC conjugation would be particularly advantageous for flow cytometry applications as it eliminates the need for secondary antibody incubation. In the validated protocol, THP-1 cells were analyzed using a fluorescently-labeled secondary antibody (DyLight®488) , suggesting that direct FITC conjugation would streamline this workflow.

What cell lines and tissues express NTSR1 and serve as appropriate positive controls?

Multiple cell lines have been validated for NTSR1 expression and can serve as positive controls:

When designing experiments with FITC-conjugated NTSR1 antibodies, these cell lines provide reliable positive controls for optimizing staining protocols.

What is the optimal protocol for flow cytometry using FITC-conjugated NTSR1 antibodies?

While the search results don't specifically address FITC-conjugated NTSR1 antibodies, a validated flow cytometry protocol using fluorescently-labeled secondary antibodies provides a methodological framework :

• Fix cells with 4% paraformaldehyde

• Block with 10% normal goat serum

• Incubate with FITC-conjugated NTSR1 antibody (approximately 1 μg per 1×10^6 cells based on validated protocol)

• Analyze using flow cytometry with appropriate filters for FITC (excitation ~495 nm, emission ~519 nm)

When adapting this protocol for FITC-conjugated antibodies, researchers should:

• Eliminate the secondary antibody incubation step

• Include appropriate controls including isotype control (FITC-conjugated rabbit IgG)

• Establish baseline with unlabeled sample

How should researchers optimize Western blot detection of NTSR1?

Search results provide detailed Western blot conditions for NTSR1 detection :

For FITC-conjugated antibodies in Western blot applications, fluorescence imaging rather than chemiluminescence would be required, potentially affecting sensitivity.

What controls are essential when using FITC-conjugated NTSR1 antibodies?

Based on validated protocols for NTSR1 detection , researchers should include:

• Unstained cells - to establish autofluorescence baseline

• Isotype control - FITC-conjugated rabbit IgG to assess non-specific binding

• Negative control - cells known not to express NTSR1

• Positive control - validated cell lines such as THP-1, MCF-7, PC-3, or DU145

• Blocking control - pre-incubation with unconjugated antibody or immunizing peptide

Search result specifically mentions using rabbit IgG (1 μg/1×10^6 cells) as an isotype control and unlabelled samples as baseline controls.

How can NTSR1 antibodies contribute to neuroscience research?

NTSR1 plays significant roles in neuronal signaling pathways. Search result indicates that "Nts can directly engage NtsR1-expressing DA neurons to modify DA signaling," highlighting its importance in dopaminergic pathways.

Researchers investigating NTSR1 in neuroscience should consider:

• Using FITC-conjugated NTSR1 antibodies to visualize receptor distribution in neuronal populations

• Combining with other neuronal markers to identify NtsR1-expressing dopaminergic neurons

• Employing the NtsR1 NEO-Cre genetic tools mentioned in search result to create conditional expression models

The cellular mechanism revealed in search result provides a foundation for understanding how neurotensin modulates dopamine signaling, making NTSR1 an important target in neuroscience research.

How are NTSR1-targeted approaches being applied in cancer research and therapeutics?

Recent research highlights significant applications of NTSR1-targeted approaches in cancer:

Search result reveals that "[177Lu]Lu-NA-ET1, an NTSR1-targeted construct" demonstrated "increased (1.9–4.4-fold) tumor retention and radiation dose delivery relative to the control." This represents a significant advancement in targeted radionuclide therapy approaches.

FITC-conjugated NTSR1 antibodies could serve as valuable tools for:

• Screening tumors for NTSR1 expression to predict therapeutic response

• Monitoring receptor dynamics following treatment

• Studying receptor internalization mechanisms that influence therapeutic efficacy

What methodological approaches can improve data quality when using FITC-conjugated NTSR1 antibodies?

To optimize experimental outcomes with FITC-conjugated NTSR1 antibodies:

• Signal amplification techniques:

  • Tyramide signal amplification for low-abundance targets

  • Multi-layer antibody approaches for enhanced sensitivity

• Microscopy optimization:

  • Confocal microscopy for improved spatial resolution

  • Deconvolution algorithms to enhance signal-to-noise ratio

  • Appropriate filter sets optimized for FITC (excitation 495 nm, emission 519 nm)

• Sample preparation:

  • Fixation with 4% paraformaldehyde as validated in flow cytometry protocol

  • Blocking with 10% normal serum to reduce non-specific binding

  • Titration of antibody concentration to determine optimal signal-to-noise ratio

What storage conditions optimize NTSR1 antibody stability and performance?

Based on search results and , optimal storage conditions include:

Search result specifically notes: "At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing."

For FITC-conjugated antibodies, additional precautions include:

• Storage in dark containers to prevent photobleaching

• Addition of preservatives (e.g., sodium azide) to prevent microbial growth

What are common technical challenges when working with NTSR1 antibodies and how can they be overcome?

Several challenges may arise when working with NTSR1 antibodies:

• Specificity concerns:

  • Validate with positive control cell lines (MCF-7, PC-3, THP-1, DU145)

  • Include appropriate negative controls and blocking controls

• Signal intensity issues:

  • For Western blot: Load adequate protein (30 μg validated)

  • For flow cytometry: Optimize antibody concentration (1 μg/1×10^6 cells validated)

• Background reduction:

  • Implement extensive washing steps (TBS with 0.1% Tween, 3× for 5 minutes each)

  • Use appropriate blocking (5% non-fat milk for Western blot, 10% normal serum for flow cytometry)

• Epitope accessibility:

  • Consider the antibody's target region (amino acids 1-70 of human NTSR1 for some antibodies)

  • Optimize fixation and permeabilization based on epitope location

How can researchers validate the specificity of NTSR1 antibodies in their experimental systems?

Comprehensive validation approaches include:

• Positive controls:

  • Use validated cell lines (MCF-7, PC-3, THP-1, DU145)

  • Expected molecular weight confirmation (43-49 kDa)

• Negative controls:

  • Cell lines without NTSR1 expression

  • Isotype control antibodies (rabbit IgG used in flow cytometry validation)

• Blocking experiments:

  • Pre-incubation with immunizing peptide

  • Competitive binding with unconjugated antibody

• Genetic validation:

  • NTSR1 knockdown/knockout controls

  • NtsR1 NEO-Cre genetic tools mentioned in search result

• Multiple detection methods:

  • Cross-validation using different techniques (Western blot, flow cytometry, ELISA)

How might advances in NTSR1-targeted therapies influence antibody-based research applications?

The development of NTSR1-targeted therapeutics creates new opportunities for antibody-based research:

• Companion diagnostics:

  • FITC-conjugated NTSR1 antibodies could serve as screening tools to identify patients likely to respond to NTSR1-targeted therapies

• Therapeutic monitoring:

  • Tracking changes in receptor expression following treatment with [177Lu]Lu-NA-ET1 or other NTSR1-targeted agents

• Mechanistic studies:

  • Investigating the "protein adducts that ranged from approximately 25–35 kDa, consistent with cysteine cathepsins" mentioned in search result

  • Exploring receptor internalization dynamics that influence therapeutic efficacy

• Multiplexed approaches:

  • Combining NTSR1 detection with other cancer biomarkers to develop comprehensive diagnostic panels

What methodological advancements might improve FITC-conjugated antibody performance in NTSR1 research?

Emerging technologies that may enhance FITC-conjugated NTSR1 antibody applications include:

• Photobleaching reduction:

  • Novel anti-fade reagents

  • Oxygen-scavenging systems for live-cell applications

• Signal enhancement:

  • FITC-tyramide signal amplification approaches

  • Quantum dot conjugation for improved photostability

• Advanced imaging methods:

  • Super-resolution microscopy to visualize receptor clustering

  • Live-cell imaging to track receptor dynamics in real-time

• Sample preparation innovations:

  • Tissue clearing techniques for three-dimensional receptor mapping

  • Optimized fixation protocols that preserve epitope accessibility while maintaining tissue architecture

By incorporating these methodological advances, researchers can maximize the utility of FITC-conjugated NTSR1 antibodies in their experimental systems.