Recombinant Mouse Protein Q300 (Hpvc2)

What is the basic structure of Mouse Protein Q300 (Hpvc2)?

Recombinant Mouse Protein Q300 (Hpvc2) is a 77-amino acid protein with UniProt ID Q02722. The full sequence is MGKCHHAHLQFHFYKFWWEGETNLFYVCVCVCVCVCVCVCTLTCMCKSGGNLGCSSSGAIHCGVFVCVLIFEPGLTM. The protein contains multiple cysteine residues that may form disulfide bonds, which are critical for maintaining its native conformation and functional properties . When expressed with an N-terminal His-tag, the recombinant form provides excellent opportunities for purification and detection in experimental settings.

How does Mouse Protein Q300 (Hpvc2) compare structurally to its human ortholog?

While the mouse Q300 (Hpvc2) protein shares significant homology with its human counterpart, researchers should note key structural differences that may impact experimental design and interpretation. Similar to how other recombinant mouse proteins like CMG-2 share 84-91% amino acid sequence homology with their human, rat, bovine, and canine orthologs , cross-species comparisons should be carefully considered when extrapolating experimental findings between models.

How can I optimize protein yield while maintaining biological activity?

Expression optimization requires balancing conditions that maximize yield while preserving biological activity. For E. coli-expressed proteins like Mouse Q300 (Hpvc2), key parameters include:

• Induction temperature (typically 18-25°C for complex proteins)

• IPTG concentration (0.1-1.0 mM)

• Expression duration (4-16 hours)

• Cell density at induction (OD600 0.6-0.8)

Researchers should conduct small-scale optimization experiments before scaling up production, as conditions that maximize yield may compromise proper folding and activity .

What is the optimal reconstitution protocol for lyophilized Mouse Protein Q300 (Hpvc2)?

For optimal reconstitution of lyophilized Mouse Protein Q300 (Hpvc2):

• Briefly centrifuge the vial before opening to bring contents to the bottom

• Reconstitute in deionized sterile water to a concentration of 0.1-1.0 mg/mL

• For long-term storage stability, add glycerol to a final concentration of 5-50% (50% is recommended)

• Aliquot to minimize freeze-thaw cycles

This methodology preserves protein structure and function while minimizing degradation during handling.

How can I prevent protein degradation during long-term storage?

To minimize degradation of Mouse Protein Q300 (Hpvc2) during storage:

• Store at -20°C/-80°C upon receipt

• Prepare multiple small aliquots to avoid repeated freeze-thaw cycles

• Store working aliquots at 4°C for up to one week

• Use Tris/PBS-based buffer with 6% Trehalose (pH 8.0) for optimal stability

• Add a carrier protein like BSA (0.1%) if using for cell culture applications

Research indicates that multiple freeze-thaw cycles significantly reduce protein activity, with each cycle potentially causing 5-15% activity loss depending on protein stability.

What are the validated applications for Recombinant Mouse Q300 (Hpvc2) in research?

Based on current research protocols, validated applications for Mouse Protein Q300 (Hpvc2) include:

• SDS-PAGE analysis for protein characterization and quality control

• Structural studies examining protein-protein interactions

• Functional assays investigating biological activity

• Development of antibodies against the target protein

Similar to approaches used with other recombinant mouse proteins, researchers can develop customized assays based on the protein's biological function and experimental objectives .

How can I design effective cell-based assays using Mouse Protein Q300 (Hpvc2)?

When designing cell-based assays with Mouse Protein Q300 (Hpvc2), consider these methodological approaches:

• Cell type selection: Use relevant mouse cell lines that express physiological binding partners

• Protein concentration optimization: Perform dose-response experiments (typically 0.1-100 ng/mL)

• Incubation conditions: Optimize temperature (37°C) and duration (30 min to 24 hours)

• Detection methods: Select appropriate antibodies or tagged versions for visualization

• Controls: Include positive controls (known interacting proteins) and negative controls

Similar proteins have been successfully used in cell assays at concentrations ranging from 0.0015-0.0023 ng/mL (ED50) for GM-CSF to 0.2-0.6 μg/mL for Syndecan-3 .

How does the His-tag affect the biological activity of Mouse Protein Q300 (Hpvc2)?

The N-terminal His-tag on Recombinant Mouse Protein Q300 (Hpvc2) facilitates purification but may impact biological activity in specific contexts. Research with similar recombinant proteins shows that:

• In most cases, N-terminal tags have minimal effect on protein folding and activity

• For some applications, tag-free versions may be preferred when the N-terminus is involved in biological interactions

• Tag position (N vs. C-terminal) can differentially affect activity depending on protein structure

• Cleavable tags with specific proteases can be used when tag-free protein is required

Researchers should validate tag effects through comparative functional assays when critical for experimental outcomes .

What strategies can improve the solubility of Mouse Protein Q300 (Hpvc2) for challenging applications?

For applications requiring high protein concentrations or challenging solution conditions:

• Buffer optimization: Screen different pH values (typically 6.5-8.5) and ionic strengths

• Additives: Include stabilizing agents such as trehalose (6%), glycerol (5-10%), or arginine (50-100 mM)

• Detergents: For hydrophobic regions, low concentrations (0.01-0.05%) of non-ionic detergents like Tween-20

• Carrier proteins: For dilute solutions, consider adding BSA (0.1-1%) to prevent adsorption to surfaces

• Temperature: Perform handling steps at 4°C when possible to reduce aggregation

These approaches have proven effective with other recombinant proteins and can be adapted for Mouse Protein Q300 (Hpvc2).

How can I verify the integrity and purity of Mouse Protein Q300 (Hpvc2) before use?

To ensure experimental reproducibility, verify protein quality using:

• SDS-PAGE: Confirm >90% purity and expected molecular weight

• Western blot: Verify identity using anti-His antibodies or protein-specific antibodies

• Mass spectrometry: Confirm exact mass and sequence coverage

• Dynamic light scattering: Assess aggregation state and homogeneity

• Activity assays: Verify biological function through application-specific assays

These quality control steps are essential for distinguishing between experimental variables and protein quality issues.

What are common troubleshooting approaches for experiments with Mouse Protein Q300 (Hpvc2)?

When experiments with Mouse Protein Q300 (Hpvc2) yield unexpected results:

• Protein activity loss:

  • Check storage conditions (avoid repeated freeze-thaw)

  • Verify buffer compatibility

  • Assess protein concentration accuracy

• Inconsistent binding:

  • Optimize binding conditions (temperature, time, pH)

  • Evaluate potential interfering substances in buffer

  • Consider lot-to-lot variability

• Aggregation issues:

  • Filter solutions before use (0.22 μm)

  • Add carrier proteins or stabilizing agents

  • Adjust protein concentration

These troubleshooting approaches are based on experimental practices documented for similar recombinant proteins .

How does handling of Mouse Protein Q300 (Hpvc2) compare to other recombinant mouse proteins?

The handling of Mouse Protein Q300 (Hpvc2) shares common principles with other recombinant mouse proteins:

This comparison highlights the importance of protein-specific handling protocols while revealing common principles across different recombinant proteins .

What functional assays can be adapted from other mouse recombinant proteins for Q300 (Hpvc2)?

Methodologies from similar proteins can be adapted for Q300 (Hpvc2) research:

• Binding assays: Similar to CMG-2 protein binding assays that use immobilized binding partners at 1.5 μg/mL

• Cell proliferation assays: Adapted from GM-CSF protocols using cell lines like FDC-P1

• Inhibition assays: Similar to AgRP C-Terminal Fragment assays measuring ED50 values (0.025-0.15 μg/mL)

• Chemotaxis assays: Borrowed from CCL22/MDC protocols measuring calcium mobilization in activated T cells

These adaptations should be validated specifically for Q300 (Hpvc2) before implementation in critical research.

What emerging technologies can advance Mouse Protein Q300 (Hpvc2) research?

Emerging technologies for recombinant protein research applicable to Mouse Protein Q300 (Hpvc2) include:

• Cryo-electron microscopy for high-resolution structural analysis

• Single-molecule biophysical techniques to study protein dynamics

• Advanced computational modeling to predict protein-protein interactions

• CRISPR-based endogenous tagging for in vivo tracking

• Cell-free expression systems for rapid production and screening

These approaches have transformed research with similar recombinant proteins and offer new avenues for understanding Q300 (Hpvc2) function .

How can in silico models enhance experimental design with Mouse Protein Q300 (Hpvc2)?

In silico approaches can significantly enhance experimental design:

• Structure prediction: Using AlphaFold or similar tools to model protein structure

• Molecular dynamics simulations: To predict protein behavior in different conditions

• Binding site prediction: To identify potential interaction partners

• Epitope mapping: For antibody development and characterization

• Stability prediction: To optimize buffer conditions and storage