Recombinant Mouse Prolactin R

Using recombinant Mouse Prolactin R in research applications allows for precise, reproducible studies of prolactin’s biological functions, including its roles in lactation, metabolism, immune regulation, and cellular differentiation. Recombinant forms provide a consistent, high-purity protein source, enabling controlled experiments in both in vitro and in vivo systems.

Key reasons to use recombinant Mouse Prolactin R include:

• Functional studies of prolactin signaling: Recombinant prolactin is biologically active and can be used to stimulate prolactin receptors in mouse cells or tissues, allowing investigation of downstream signaling pathways and physiological effects.
• Metabolic and endocrine research: Prolactin influences metabolic homeostasis, including effects on the pancreas, liver, adipose tissue, and hypothalamus. Recombinant prolactin has been shown to improve metabolic profiles in rodent models of diabetes and obesity, making it valuable for metabolic disease studies.
• Lactation and reproductive biology: Prolactin is essential for lactation and mammary gland development. Recombinant prolactin can restore lactational performance in animal models where endogenous prolactin is suppressed, supporting studies on lactation physiology and potential therapeutic interventions.
• Immunological and vascular studies: Prolactin has immune-regulatory functions and influences vascular growth. Recombinant prolactin enables controlled experiments to dissect these roles in mouse models.
• Cellular differentiation assays: In vitro, recombinant mouse prolactin modulates gene expression related to adipocyte differentiation, supporting studies on cell fate and tissue development.
• Consistency and reproducibility: Recombinant proteins are produced under defined conditions, ensuring batch-to-batch consistency and eliminating variability associated with pituitary extracts or serum-derived proteins.

Additional advantages:

• High purity and activity: Modern recombinant production methods yield prolactin with high purity and confirmed biological activity, as validated by cell-based assays.
• Cost-effectiveness and scalability: Recombinant production in systems like E. coli is efficient and scalable, facilitating both small-scale laboratory research and larger preclinical studies.

In summary, recombinant Mouse Prolactin R is a versatile tool for dissecting prolactin’s diverse physiological roles in mouse models, supporting research in endocrinology, metabolism, immunology, and developmental biology with high specificity and reproducibility.

Yes, recombinant mouse prolactin (PRL) can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for your specific assay system. Recombinant PRL is commonly used as a standard in mouse prolactin ELISA kits and is suitable for generating standard curves to quantify PRL in biological samples.

Key considerations and best practices:

• Immunoreactivity: Most commercial mouse PRL ELISA kits are designed to detect both natural and recombinant mouse PRL, and the standard curves are often generated using recombinant PRL. However, it is essential to confirm that the recombinant PRL you use is recognized equivalently by the antibodies in your specific ELISA system.
• Standard Preparation: Prepare the recombinant PRL standard according to the recommended protocol, ensuring accurate dilution to match the assay’s dynamic range (typically from low pg/mL to ng/mL concentrations).
• Matrix Effects: Dilute the recombinant standard in the same buffer or matrix as your samples (e.g., assay diluent, serum, or plasma) to minimize matrix effects and ensure accurate quantification.
• Value Assignment: The mass value on the recombinant protein vial may not always correspond exactly to the immunoreactive mass detected by the ELISA, due to differences in protein folding, post-translational modifications, or lot-to-lot variability. It is best practice to assign the standard’s value based on its performance in the ELISA, not solely on the mass stated on the vial.
• Validation: Validate the use of your recombinant PRL standard by running a standard curve and comparing it to any kit-provided standards or by spiking known amounts into sample matrices to assess recovery and linearity.

Summary Table: Use of Recombinant Mouse Prolactin as ELISA Standard

In conclusion: Recombinant mouse prolactin is widely accepted and scientifically appropriate as a standard for ELISA quantification, provided you validate its performance in your specific assay context.

Recombinant Mouse Prolactin (mPRL) has been validated for several applications in published research, primarily focusing on its biological activity and functional effects in both in vitro and in vivo settings. Key applications include:

1. In Vivo Assays:

   • Used in animal models to study the effects of prolactin on physiological processes such as lactation, neurogenesis, and metabolic adaptations. For example, recombinant mouse prolactin has been administered in vivo to assess its role in stimulating precursor cells in the adult mouse hippocampus and in models of lactation and galactopoiesis.

2. Bioassays:

   • Employed in bioassays to evaluate prolactin receptor (PRLR) signaling, including cell proliferation assays using murine Ba/F3 cells stably expressing PRLR. These assays measure the ability of recombinant prolactin to activate downstream signaling pathways.

3. Cell-Based Functional Studies:

   • Applied in cell culture systems to investigate the effects of prolactin on gene expression and cellular differentiation. For instance, recombinant mouse prolactin has been used to study adipocyte differentiation in 3T3-L1 cells, where it modulates the expression of key transcription factors such as PPARγ, C/EBPβ, and FAS.

4. Pharmacokinetic and Pharmacodynamic Studies:

   • Engineered variants of recombinant prolactin (e.g., Prolactin-XL) have been tested for their persistence, half-life, and efficacy in mouse models, including studies on drug exposure and clearance rates.

5. Cytotoxicity and Cell Viability Assays:

   • Evaluated for cytotoxicity in cell lines such as 3T3-L1, confirming that recombinant mouse prolactin does not adversely affect cell viability at biologically relevant concentrations.

These applications demonstrate that recombinant mouse prolactin is a versatile tool for studying prolactin biology, receptor signaling, and its roles in development, metabolism, and disease.

Reconstitution Protocol

Recombinant Mouse Prolactin R protein is typically supplied as a lyophilized powder formulated in phosphate-buffered saline (PBS) with 0.2 µm filtration. Before reconstitution, briefly spin the vial and allow the contents to settle at the bottom to ensure complete dissolution.

The recommended reconstitution concentration is 0.5–1.0 mg/mL using sterile deionized water. For formulations containing carrier proteins, reconstitute at 100 µg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA). Allow the vial to reconstitute for 15–30 minutes at room temperature with gentle agitation, avoiding vigorous shaking that can cause foaming and protein denaturation.

Preparation for Cell Culture

Carrier Protein Addition: For long-term storage and stability during cell culture experiments, add a carrier protein such as 0.1% BSA or human serum albumin (HSA) to the reconstituted solution. This prevents non-specific adsorption to container surfaces and maintains protein integrity.

Handling Considerations: During reconstitution, the protein may appear as a film at the bottom of the vial. Gently mix the vial after adding the diluent rather than vigorous mixing. Avoid repeated freeze-thaw cycles by using a manual defrost freezer and storing reconstituted aliquots in working portions.

Storage Stability: Reconstituted prolactin remains stable for at least 3 months when stored in working aliquots with carrier protein at –20°C. Store the original lyophilized vial at the temperature specified in the product documentation immediately upon receipt.

Protein Characteristics

The recombinant mouse prolactin receptor protein has a calculated molecular mass of approximately 27 kDa and is typically expressed in mammalian systems with an N-terminal histidine tag. The protein is purified to greater than 95% purity as determined by SDS-PAGE under reducing conditions. For cell-based assays, typical effective concentrations range from 0.25–1 ng/mL depending on the specific application.