Recombinant Human HYAL4 Protein: Handling & Application Protocol

Recombinant Human HYAL4 Protein is a member of the hyaluronidase family produced recombinantly for use in biochemical, enzymatic, and cell-based research applications requiring high-purity, functionally active protein.

Product Name

Recombinant Human HYAL4 Protein

SKU

rp377630

Protein Family

Hyaluronidase (HYAL) family

Source

Recombinant expression system

Recommended Storage

-20°C or -80°C; avoid repeated freeze-thaw cycles

Application

Biochemical assays, enzymatic activity studies, cell-based research

Overview of Recombinant Human HYAL4 Protein

HYAL4 (Hyaluronidase-4) is a member of the hyaluronidase superfamily, enzymes that degrade hyaluronic acid (HA) and other glycosaminoglycans in the extracellular matrix. The HYAL gene family includes HYAL1–HYAL4 and SPAM1, each with distinct tissue distribution, substrate specificity, and functional roles. HYAL4 has been reported to preferentially act on chondroitin sulfate rather than hyaluronic acid, distinguishing it functionally from other family members.

Recombinant Human HYAL4 Protein (SKU: rp377630) from Aladdin Scientific is produced using recombinant technology to deliver a well-defined, research-grade protein suitable for in vitro studies. It is intended exclusively for research use and is not approved for diagnostic or therapeutic applications.

Reagents, Equipment, and Safety Precautions

Required Reagents:

• Recombinant Human HYAL4 Protein (rp377630)
• Sterile, ultrapure water or manufacturer-recommended reconstitution buffer (e.g., PBS, pH 7.4)
• Carrier protein (e.g., 0.1% BSA) if required for stabilization
• Appropriate assay buffer matched to downstream application

Equipment:

• Microcentrifuge
• Low-protein-binding microtubes
• Ice bucket or cooling block
• Calibrated pipettes

Safety: Handle the protein following standard laboratory biosafety guidelines for recombinant proteins. Wear appropriate PPE including gloves and eye protection. Refer to the Safety Data Sheet (SDS) before use. The protein is not known to present specific biohazardous risk at research-use quantities, but standard precautions apply.

Reconstitution and Storage Protocol

Step 1 – Preparation: Allow the vial to equilibrate to room temperature for 10–15 minutes before opening. Briefly centrifuge the vial (1,000 × g, 30 seconds) to collect the lyophilized material at the bottom.

Step 2 – Reconstitution: Reconstitute the lyophilized protein by adding the appropriate volume of sterile ultrapure water or the buffer recommended in the product datasheet to achieve the desired stock concentration. Gently pipette up and down or swirl carefully; do not vortex. Allow complete dissolution for 5–10 minutes on ice.

Step 3 – Carrier Protein Addition (optional): To enhance stability, especially for dilute working solutions, consider adding BSA to a final concentration of 0.1% (w/v). This is particularly recommended when aliquoting protein at low concentrations.

Step 4 – Aliquoting: Divide the reconstituted protein into single-use aliquots using low-protein-binding microtubes to minimize repeated freeze-thaw cycles and adsorptive losses.

Step 5 – Storage: Store aliquots at -20°C for short-term use (up to 1 month) or at -80°C for long-term storage. Avoid repeated freeze-thaw cycles, as these can reduce protein activity and integrity. Once thawed, keep on ice and use promptly.

Note: Always consult the official product datasheet for lot-specific reconstitution volumes and storage recommendations.

Experimental Applications and Usage Guidelines

Recombinant Human HYAL4 Protein is suitable for a range of in vitro research applications:

• Enzymatic Activity Assays: HYAL4 can be used to study glycosaminoglycan (GAG) degradation, particularly chondroitin sulfate cleavage, using fluorometric or colorimetric substrate assays.
• Biochemical Interaction Studies: The protein can serve as a binding partner or tool reagent in pull-down assays, surface plasmon resonance (SPR), or ELISA-based interaction studies investigating extracellular matrix remodeling pathways.
• Cell-Based Assays: Exogenously added HYAL4 protein may be used to modify pericellular glycosaminoglycan matrices in cell culture models to examine effects on cell signaling, migration, or proliferation.
• Antibody Validation: Recombinant HYAL4 can serve as a positive control antigen in Western blot, ELISA, or immunoprecipitation experiments validating HYAL4-specific antibodies.

Determine the optimal working concentration empirically for each assay system. Typical starting concentrations for enzymatic assays should be titrated based on specific activity data provided in the product datasheet.

Troubleshooting and Quality Control

No or Low Enzymatic Activity:

• Verify that the protein was reconstituted correctly and that the recommended buffer and pH were used.
• Confirm that the working concentration is within the expected active range per the datasheet.
• Check that the substrate used is appropriate for HYAL4 (chondroitin sulfate preferred over hyaluronic acid).
• Ensure the protein has not undergone excessive freeze-thaw cycles.

Protein Precipitation or Aggregation:

• Avoid vortexing during reconstitution.
• Include carrier protein (BSA) in dilute solutions.
• Use low-protein-binding tubes and pipette tips.

Quality Control: Aladdin Scientific's recombinant proteins undergo internal QC testing. Confirm purity, expected molecular weight by SDS-PAGE, and biological activity data from the certificate of analysis (CoA) included with each lot. Contact Aladdin Scientific technical support if results are inconsistent with CoA specifications.

Frequently asked questions

What buffer should I use to reconstitute Recombinant Human HYAL4 Protein?

The recommended reconstitution buffer is typically sterile ultrapure water or PBS (pH 7.4), as specified in the product datasheet for lot rp377630. Always consult the CoA provided with your specific lot for precise reconstitution instructions.

How many freeze-thaw cycles can Recombinant Human HYAL4 Protein tolerate?

It is generally recommended to limit freeze-thaw cycles to a minimum, ideally no more than 2–3 cycles, to preserve protein activity and structural integrity. Single-use aliquoting upon first reconstitution is strongly advised.

Is Recombinant Human HYAL4 Protein suitable for in vivo studies?

No. Recombinant Human HYAL4 Protein (rp377630) from Aladdin Scientific is intended for research use only and is not suitable or approved for in vivo, diagnostic, or therapeutic applications.

What is the preferred substrate for HYAL4 enzymatic activity assays?

HYAL4 has been reported to preferentially degrade chondroitin sulfate rather than hyaluronic acid, which distinguishes it from other hyaluronidase family members such as HYAL1. Use chondroitin sulfate-based substrates for optimal enzymatic activity detection.

How should I verify the quality of my HYAL4 protein lot?

Review the Certificate of Analysis (CoA) accompanying your lot for purity assessment by SDS-PAGE, expected molecular weight, and biological activity data. If any discrepancy is noted between your experimental results and the CoA, contact Aladdin Scientific technical support for assistance.

References

1. Csóka AB, Scherer SW, Stern R. Expression analysis of six paralogous human hyaluronidase genes clustered on chromosomes 3p21 and 7q31. Genomics. 1999;60(3):356-361.
2. Stern R, Jedrzejas MJ. Hyaluronidases: their genomics, structures, and mechanisms of action. Chemical Reviews. 2006;106(3):818-839.
3. Kaneiwa T, Mizumoto S, Sugahara K, Yamada S. Identification of human hyaluronidase-4 as a novel chondroitin sulfate hydrolase that preferentially cleaves the galactosaminidic linkage in the trisulfated tetrasaccharide sequence. Glycobiology. 2010;20(3):300-309.

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.