Recombinant Rat Prolactin Receptor

Using Recombinant Rat Prolactin Receptor (PRLR) in research applications enables precise investigation of prolactin signaling, receptor-ligand interactions, and downstream biological effects in rat models, which are highly relevant for studies in endocrinology, oncology, neurobiology, and immunology.

Key scientific advantages and applications include:

• Receptor-Ligand Binding Studies: Recombinant PRLR allows for controlled in vitro assays to characterize prolactin binding kinetics, affinity, and specificity, facilitating mechanistic studies of hormone-receptor interactions.
• Signal Transduction Analysis: Using recombinant PRLR enables dissection of downstream signaling pathways activated by prolactin, such as JAK-STAT, MAPK, and PI3K, which are implicated in cell proliferation, differentiation, and survival.
• Cancer Research: PRLR is expressed in various tumor types, including glioblastoma and breast cancer. Recombinant PRLR can be used to model receptor activation, study chemoresistance mechanisms, and evaluate potential therapeutic antagonists or inhibitors targeting the prolactin pathway.
• Immunological Studies: PRLR is present on immune cells, and recombinant receptor can be used to investigate prolactin’s role in immune modulation, cytokine regulation, and T-cell function.
• Drug Screening and Therapeutic Development: Recombinant PRLR provides a standardized platform for screening small molecules, antibodies, or peptides that modulate prolactin signaling, supporting preclinical drug development.
• ELISA Standards and Controls: Recombinant PRLR is suitable as a standard or control in quantitative assays such as ELISA, ensuring reproducibility and accuracy in measuring endogenous receptor levels.
• Species-Specific Research: Rat PRLR is essential for studies where species-specific receptor biology is critical, such as comparative endocrinology or translational research using rat models.

Best practices for use:

• Employ recombinant PRLR in cell-based assays, receptor binding studies, and biochemical analyses where precise control over receptor concentration and purity is required.
• Validate recombinant receptor activity and specificity in your assay system, especially when comparing to endogenous receptor responses.

In summary, Recombinant Rat Prolactin Receptor is a versatile tool for elucidating prolactin biology, modeling disease mechanisms, and advancing therapeutic research in rat systems.

You can use recombinant Rat Prolactin Receptor as a standard for quantification or calibration in ELISA assays, provided it is well-characterized, pure, and its concentration is accurately determined.

For ELISA quantification, the standard should:

• Be a purified or recombinant protein with a known concentration, ideally verified by an independent method such as HPLC or spectrophotometry.
• Be reconstituted and diluted according to best practices, using serial dilutions to cover the expected range of your samples.
• Match the epitope recognized by the assay antibodies; recombinant proteins may differ in folding or post-translational modifications compared to native proteins, which can affect antibody binding and quantification accuracy.

Key considerations:

• Ensure the recombinant protein is biologically and immunologically equivalent to the native rat prolactin receptor, especially regarding antibody recognition in your ELISA format.
• If your recombinant protein includes tags or differs in structure, verify that the ELISA antibodies can detect it with the same efficiency as the native protein.
• Large dilution steps from stock concentrations (often μg/mL to pg/mL) can introduce error; use precise pipetting and validated protocols to minimize inaccuracies.
• When using recombinant proteins as standards, a recovery of ±25% compared to the stated amount is typical in ELISA quantification, due to inherent assay variability.
• For best practice, include a spike control (standard diluted in sample matrix) to confirm that matrix effects do not interfere with quantification.

Protocol tips:

• Prepare a standard curve using serial dilutions of the recombinant protein, covering the expected concentration range of your samples.
• Validate the standard curve by back-calculating known concentrations and ensuring the R² value is ≥0.99 for confidence in fit.
• If possible, compare results using both recombinant and endogenous standards to confirm assay accuracy.

In summary, recombinant Rat Prolactin Receptor is suitable as an ELISA standard if it is pure, accurately quantified, and immunologically compatible with your assay antibodies. Always validate its performance in your specific assay context.

The Recombinant Rat Prolactin Receptor has been validated for several key applications in published research, primarily focusing on its role in receptor-ligand interaction studies and downstream signaling pathways. Based on the available literature and product documentation:

1. Receptor Binding Assays: The recombinant rat prolactin receptor is used to characterize prolactin receptor interactions and binding kinetics with prolactin and other ligands. This is essential for understanding the molecular mechanisms of prolactin signaling and for screening potential agonists or antagonists.

2. Signal Transduction Studies: The receptor participates in major signaling pathways such as the JAK2-STAT5 and PI3K-AKT pathways. These pathways are critical for mediating the biological effects of prolactin, including cell proliferation, survival, and metabolism. Studies have used the receptor to investigate these signaling cascades in vitro.

3. Cancer Research: Prolactin receptor signaling is upregulated in various hormone-dependent cancers, including breast, prostate, ovarian, and endometrial cancers. The recombinant receptor has been used to study the effects of prolactin receptor antagonists and to explore the therapeutic potential of targeting PRLR in cancer treatment.

4. ELISA and Western Blotting: The receptor is validated for use in enzyme-linked immunosorbent assays (ELISA) and western blotting, allowing for the detection and quantification of prolactin receptor expression and its interactions.

5. Functional Assays: The receptor is used in functional assays to assess the biological activity of prolactin and its analogs, including their effects on cell proliferation, survival, and migration.

These applications highlight the versatility of the Recombinant Rat Prolactin Receptor in both basic research and translational studies, particularly in the fields of endocrinology, oncology, and molecular biology.

To reconstitute and prepare Recombinant Rat Prolactin Receptor (PRLR) protein for cell culture experiments, follow these technical steps:

• Reconstitution:

  • Use sterile, deionized water or PBS to reconstitute the lyophilized PRLR protein.
  • Recommended concentration for reconstitution is 0.1–1.0 mg/mL.
  • For some preparations, do not reconstitute below 100 μg/mL to maintain protein stability.
  • Gently pipette the solution down the sides of the vial; do not vortex.
  • Allow the protein to dissolve completely at room temperature for 10–30 minutes. If particulate remains, gentle mixing for up to 2 hours may be required.

• Aliquoting and Storage:

  • After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles.
  • For long-term storage, add a carrier protein such as 0.1% BSA or HSA to prevent adsorption and degradation.
  • Store aliquots at –80°C for long-term stability; short-term storage (days) can be at 4–8°C.
  • Avoid repeated freeze-thaw cycles to preserve protein integrity.

• Preparation for Cell Culture:

  • Before use, thaw aliquots on ice and dilute to the desired working concentration in cell culture medium or appropriate buffer.
  • If required, supplement with glycerol (5–50%) for additional stability, especially for functional assays.
  • Filter sterilize the final solution if necessary, using a 0.2 μm filter to ensure sterility for cell culture applications.

• General Best Practices:

  • Centrifuge the vial briefly before opening to collect the powder at the bottom.
  • Equilibrate all reagents and protein solutions to room temperature before use.
  • Quantify protein concentration using a suitable assay (e.g., BCA or Bradford) after reconstitution if precise dosing is required.

Summary Table: Recombinant Rat PRLR Protein Preparation

These protocols ensure protein stability, activity, and sterility for cell culture experiments. Always consult the specific product datasheet for any unique requirements related to your recombinant PRLR preparation.