Recombinant Mouse PF-4

Recombinant Mouse PF-4 (Platelet Factor 4, CXCL4) is used in research applications to study its roles in inflammation, angiogenesis, hematopoiesis, and immune regulation, as well as to develop and validate assays and investigate disease mechanisms in mouse models.

Key reasons to use recombinant Mouse PF-4 in your research:

• Defined Activity and Consistency: Recombinant PF-4 provides a highly purified, well-characterized protein with consistent biological activity, essential for reproducible results in functional assays and mechanistic studies.
• Angiogenesis and Endothelial Cell Studies: Mouse PF-4 is a known inhibitor of angiogenesis. For example, recombinant Mouse PF-4 has been shown to inhibit FGF-basic-dependent proliferation of human umbilical vein endothelial cells (HUVECs), making it valuable for studying vascular biology and tumor angiogenesis.
• Immunology and Inflammation: PF-4 is a CXC chemokine involved in modulating immune cell recruitment and inflammatory responses. Using recombinant protein allows for controlled studies on chemotaxis, cytokine signaling, and immune cell interactions in vitro and in vivo.
• Assay Development and Standardization: Recombinant PF-4 is commonly used as a standard in ELISA and other immunoassays, enabling quantification of PF-4 in biological samples and validation of antibody specificity.
• Hematopoiesis and Platelet Biology: PF-4 is released from platelets and regulates megakaryopoiesis and hematopoietic stem cell function. Recombinant PF-4 enables detailed studies of these processes in mouse models, which are relevant for thrombosis, hemostasis, and bone marrow research.
• Disease Modeling: Mouse PF-4 is implicated in various disease models, including atherosclerosis, cancer, and autoimmune disorders. Recombinant protein allows for precise dosing and mechanistic dissection in preclinical studies.

Additional advantages of recombinant proteins:

• High Purity and Activity: Recombinant Mouse PF-4 is typically purified to >97% and retains its biological activity, ensuring experimental reliability.
• Tag-Free or Tagged Options: Availability of tag-free or His-tagged versions allows flexibility for different assay formats and purification needs.
• Batch-to-Batch Consistency: Recombinant production ensures minimal variability between lots, which is critical for quantitative and comparative studies.

In summary, using recombinant Mouse PF-4 enables precise, reproducible, and mechanistically informative studies in mouse systems, supporting a wide range of research applications in vascular biology, immunology, hematology, and disease modeling.

Yes, recombinant mouse PF-4 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated and matched to the assay system. Recombinant PF-4 is commonly used as a standard in commercial mouse PF-4 ELISA kits, and dose-response curves generated with recombinant PF-4 are typically parallel to those obtained with natural PF-4, supporting its suitability for quantification.

Key considerations and supporting details:

• Validation: Commercial ELISA kits for mouse PF-4 routinely use recombinant PF-4 as the standard. These kits demonstrate that recombinant PF-4 yields parallel standard curves to natural PF-4, indicating comparable immunoreactivity and quantification accuracy.
• Purity and Quantification: For accurate calibration, the recombinant PF-4 standard should be highly purified, and its concentration must be precisely determined, typically by HPLC or absorbance-based protein quantification methods.
• Matrix Compatibility: Recovery and linearity studies in ELISA kits show that recombinant PF-4 can be spiked into various sample matrices (e.g., plasma, serum, cell culture supernatants) with high recovery rates, further supporting its use as a standard.
• Preparation: When preparing your own standard curve, dilute the recombinant PF-4 in the same buffer or diluent used for your samples to minimize matrix effects and ensure consistency.

Best Practices:

• Use recombinant PF-4 that matches the species and isoform of your target analyte.
• Validate the standard curve by running serial dilutions and confirming linearity and parallelism with endogenous PF-4 in your sample matrix.
• Store and handle recombinant PF-4 according to manufacturer or protocol recommendations to maintain stability and activity.

Summary Table: Recombinant PF-4 as ELISA Standard

In conclusion, recombinant mouse PF-4 is widely accepted and scientifically validated as a standard for ELISA quantification and calibration, provided it is properly prepared and matched to your assay system.

Recombinant Mouse PF-4 (CXCL4) has been validated in published research for several applications, including functional cell-based assays, immunoassays (such as ELISA), SDS-PAGE, and as a tool in preclinical and comparative immunology studies.

Key validated applications and supporting details:

• Functional Cell-Based Assays:
  Recombinant Mouse PF-4 has been shown to inhibit FGF basic-dependent proliferation of human umbilical vein endothelial cells (HUVECs), demonstrating its use in angiogenesis and cell proliferation assays. It has also been used to modulate inflammatory signaling and immune cell gene expression in vivo, including RNA-seq analysis of microglia and behavioral testing in mouse models of aging and cognition.

• Immunoassays (ELISA):
  The protein is validated as a standard for sandwich ELISA (sELISA), enabling quantification of PF-4 in biological samples.

• SDS-PAGE and Western Blot:
  Recombinant Mouse PF-4 is suitable for purity assessment and molecular weight confirmation by SDS-PAGE. It is also used as a positive control or immunogen in Western blotting.

• Preclinical and Comparative Immunology Research:
  PF-4 is used as a reagent in mouse model studies, comparative immunology, and preclinical therapeutic evaluations, particularly for its roles in inflammation, immune modulation, and cognitive function.

• In Vivo Functional Studies:
  Systemic administration of recombinant PF-4 in mice has been validated for studies on neuroinflammation, microglial gene expression, and cognitive performance, including behavioral assays such as novel object recognition and Y maze testing.

• Protein-Protein Interaction Studies:
  PF-4 has been used to study interactions with viral vectors (e.g., adenovirus), relevant to vaccine research and immunogenicity.

Summary Table of Validated Applications

Note:
The specific validation of each application may depend on the recombinant PF-4 preparation and the experimental context. Published studies have demonstrated its biological activity in both in vitro and in vivo systems, supporting its use in a wide range of research applications.

To reconstitute and prepare Recombinant Mouse PF-4 (CXCL4) protein for cell culture experiments, follow these best-practice steps:

1. Equilibrate and Centrifuge the Vial

   • Allow the lyophilized protein vial and your reconstitution buffer (typically sterile, deionized water) to reach room temperature.
   • Briefly centrifuge the vial (20–30 seconds) to collect all powder at the bottom and minimize loss.

2. Reconstitution

   • Add sterile, deionized water to achieve a final concentration of 0.1–1.0 mg/mL. Do not reconstitute below 0.1 mg/mL to avoid protein instability.
   • Gently mix by pipetting up and down or swirling. Do not vortex or shake vigorously, as this can denature the protein.
   • If the protein does not dissolve immediately, allow it to sit at room temperature for 15–30 minutes with gentle agitation. If visible flakes remain, extend gentle mixing up to 2 hours.

3. Carrier Protein Addition (Optional but Recommended)

   • For enhanced stability, especially at low concentrations or for storage, add a carrier protein such as 0.1% BSA or 10% FBS to the solution.
   • For long-term storage, adding 5–50% glycerol is also recommended.

4. Aliquot and Storage

   • Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can degrade the protein.
   • Store aliquots at –20°C to –80°C for long-term storage, or at 2–8°C for up to 1 month if used soon.

5. Sterility

   • Ensure all solutions and containers are sterile to prevent contamination in cell culture applications.

6. Working Solution Preparation

   • Before adding to cell cultures, dilute the stock solution to the desired working concentration using sterile cell culture medium or buffer containing carrier protein if needed.

Summary Table: Key Steps for Recombinant Mouse PF-4 Reconstitution

Additional Notes:

• Avoid repeated freeze-thaw cycles to maintain protein integrity.
• Confirm protein concentration and integrity if needed by SDS-PAGE or other analytical methods.
• Always consult the specific product datasheet or Certificate of Analysis for any lot-specific instructions.

These guidelines are based on standard protocols for recombinant chemokines and PF-4 proteins and are suitable for most cell culture applications.