PTGR2 Antibody, HRP conjugated

What is PTGR2 and what biological functions does it serve?

PTGR2 (Prostaglandin Reductase 2) functions as a 15-oxo-prostaglandin 13-reductase that catalyzes 15-keto-PGE2 (an endogenous PPARγ ligand) into 13,14-dihydro-15-keto-PGE2. The enzyme acts primarily on 15-keto-PGE1, 15-keto-PGE2, 15-keto-PGE1-alpha, and 15-keto-PGE2-alpha, with highest catalytic activity toward 15-keto-PGE2 .

PTGR2 has demonstrated oncogenic properties in both gastric and pancreatic cancers. Research indicates that overexpression of PTGR2 represses the transcriptional activity of PPARG and inhibits adipocyte differentiation. From a clinical perspective, PTGR2 has been found to modulate reactive oxygen species (ROS)-mediated cell death and tumor transformation .

How do I select the appropriate application for PTGR2 antibody, HRP conjugated?

When selecting applications for PTGR2 antibody with HRP conjugation, consider the following methodological approach:

• Determine your detection requirements: HRP-conjugated antibodies are particularly well-suited for:

  • ELISA-based quantification (direct detection without secondary antibody)

  • Immunohistochemistry with enhanced sensitivity

  • Western blotting applications requiring direct detection

• Validate specificity: Confirm the antibody's reactivity with human samples in your specific application. Available data shows that the antibody effectively recognizes PTGR2 in human cell lines including HEK-293, HepG2, and HeLa .

• Consider dilution factors: For Western blot applications, successful results have been demonstrated at 1/500 dilution with human cell lysates. Optimization may be required for your specific experimental conditions .

What are the optimal storage and handling conditions for maintaining antibody integrity?

To preserve PTGR2 antibody activity and stability:

• Upon receipt, store at -20°C or -80°C to prevent degradation .

• Avoid repeated freeze-thaw cycles which can denature antibody proteins and reduce binding efficiency.

• The antibody is supplied in a preservative solution containing 0.03% Proclin 300, 50% Glycerol, and 0.01M PBS (pH 7.4), which helps maintain stability during storage .

• When working with the antibody, maintain cold chain practices by keeping it on ice during experiments.

How do I optimize ELISA protocols using PTGR2 antibody, HRP conjugated?

When optimizing ELISA protocols with PTGR2 antibody:

• Detection range consideration: Commercial PTGR2 ELISA systems demonstrate a detection range of 0.156-10 ng/ml with a sensitivity of 0.059 ng/ml. Use this as a benchmark when establishing your detection parameters .

• Sandwich ELISA methodology: For maximum sensitivity and specificity:

  • Coat plates with a capture antibody specific to a different epitope than your HRP-conjugated antibody

  • Add samples and standards in duplicate or triplicate

  • Add the HRP-conjugated PTGR2 antibody (working dilution requires optimization)

  • Wash thoroughly between steps to reduce background

  • Develop with TMB substrate and measure absorbance at 450nm

• Sample preparation: PTGR2 antibody has been validated for:

  • Human serum

  • Plasma (specifically validated for citrated/EDTA plasma)

  • Cell culture supernatants

What controls should I include when using PTGR2 antibody in Western blot applications?

For robust Western blot experiments using PTGR2 antibody:

• Positive controls: Include lysates from cell lines known to express PTGR2, such as:

  • HEK-293 (human embryonic kidney cells)

  • HepG2 (human liver hepatocellular carcinoma)

  • HeLa (human cervical adenocarcinoma)

• Negative controls:

  • Primary antibody omission control

  • Non-specific rabbit IgG at equivalent concentration

  • Lysates from cell lines with confirmed low PTGR2 expression

• Loading controls: Include antibodies against housekeeping proteins (e.g., GAPDH, β-actin) to normalize protein loading across lanes.

How can I troubleshoot high background issues in immunoassays using HRP-conjugated antibodies?

When encountering high background with PTGR2 HRP-conjugated antibody:

• Increase washing stringency: Extend washing time and number of washes between incubation steps using fresh buffer.

• Optimize antibody dilution: Titrate the HRP-conjugated antibody concentration to determine optimal signal-to-noise ratio. Start with manufacturer-recommended dilution (e.g., 1/500 for Western blot) and adjust as needed .

• Block more effectively: Use 5% non-fat dry milk or BSA in TBST buffer for Western blot applications. For ELISA, commercially available blocking reagents can reduce non-specific binding.

• Reduce substrate exposure time: Shorten the development time with TMB or other HRP substrates to minimize background signal development.

How can PTGR2 antibody be used to investigate its role in oncogenic processes?

Based on established oncogenic links, PTGR2 antibody can be employed to:

• Evaluate PTGR2 expression in tumor tissues: Immunohistochemistry studies have demonstrated that 85.5% of pancreatic ductal adenocarcinoma tissues stain positive for PTGR2 expression while adjacent normal tissues show minimal staining .

• Investigate ROS regulation: Design experiments to examine how PTGR2 expression levels correlate with ROS production in cancer cells by:

  • Using the antibody to confirm PTGR2 silencing efficiency in knockdown experiments

  • Quantifying PTGR2 protein levels before measuring cellular ROS

  • Correlating PTGR2 expression with cell proliferation and death markers

• Study mechanistic pathways: Research indicates PTGR2 silencing suppresses expressions of solute carrier family 7 member 11 (xCT). Design experiments to investigate this and other downstream regulatory pathways using the antibody as a confirmatory tool .

What considerations should be made when designing experiments to analyze PTGR2 expression across different cancer types?

When investigating PTGR2 across cancer types:

• Tissue-specific normalization:

  • Establish appropriate baseline expression levels for each tissue type

  • Include paired normal-tumor samples when possible

  • Consider that 85.5% of pancreatic ductal adenocarcinomas show PTGR2 positivity, which can serve as a reference point

• Quantitative analysis method selection:

  • For sensitive quantitation in research samples, ELISA with a detection sensitivity of 0.059 ng/ml can be employed

  • For spatial distribution analysis, immunohistochemistry with appropriate controls is recommended

  • For protein size confirmation and semi-quantitative analysis, Western blotting provides valuable insights

• Clinical correlation consideration:

  • Current research suggests PTGR2 stain intensity isn't significantly associated with differentiation status or clinical stage in pancreatic ductal adenocarcinoma patients

  • Design experiments to investigate whether this holds true across other cancer types

What are the detailed specifications of commercially available PTGR2 antibody, HRP conjugated?

The following specifications are important for experimental planning:

How can I validate the specificity of the PTGR2 antibody in my experimental system?

To confirm antibody specificity:

• Peptide competition assay: Pre-incubate the antibody with excess immunogen peptide (PTGR2 aa 206-282) before application to your samples. Loss of signal confirms specificity.

• Knockdown validation: Compare antibody reactivity in:

  • Wild-type cells/tissues

  • PTGR2 knockdown cells (siRNA or CRISPR)

  • PTGR2 overexpression systems

• Cross-reactivity testing: Test the antibody against recombinant proteins with similar structural domains to PTGR2, particularly other zinc-binding alcohol dehydrogenase domain-containing proteins.

• Western blot migration validation: Confirm that the detected protein migrates at the expected molecular weight for PTGR2.

How can PTGR2 antibody be utilized in prostaglandin metabolism research?

The antibody can facilitate investigations into prostaglandin metabolism through:

• Enzyme activity correlation studies:

  • Quantify PTGR2 protein levels using the antibody

  • Correlate with 15-oxo-prostaglandin 13-reductase activity measurements

  • Investigate how PTGR2 levels affect the conversion of 15-keto-PGE2 to 13,14-dihydro-15-keto-PGE2

• PPARγ signaling research:

  • Use the antibody to monitor PTGR2 expression levels

  • Correlate with PPARγ transcriptional activity

  • Investigate the relationship between PTGR2 expression and adipocyte differentiation

• Biomarker development:

  • Evaluate PTGR2 as a potential biomarker for cancers by quantifying expression in patient samples

  • Correlate expression levels with clinical outcomes and treatment responses

What methodological approaches can be employed to study the relationship between PTGR2 and reactive oxygen species in cancer progression?

To investigate PTGR2's relationship with ROS in cancer:

• Sequential analysis approach:

  • First, confirm PTGR2 expression levels using the antibody in various cancer cell models

  • Then measure intracellular ROS using fluorescent probes

  • Finally, correlate PTGR2 expression with ROS levels, cell proliferation, and cell death markers

• Mechanism exploration methodology:

  • Use the antibody to confirm successful manipulation of PTGR2 expression (overexpression or silencing)

  • Monitor changes in the expression of ROS-regulating genes (e.g., xCT)

  • Measure the direct effects on 15-keto-PGE2 levels, which may mediate the observed phenotypes

• Therapeutic intervention testing:

  • Evaluate how modulating PTGR2 expression affects sensitivity to ROS-inducing cancer therapies

  • Use the antibody to confirm target engagement in preclinical models