Recombinant Human C6orf163 Protein: Handling & Usage Protocol

Recombinant Human C6orf163 Protein is a research-grade recombinant protein requiring careful reconstitution, aliquoting, and storage to maintain full biological activity and ensure reproducible results across downstream assays.

Product Name

Recombinant Human C6orf163 Protein

SKU

rp376452

Species Origin

Human (Homo sapiens)

Protein Type

Recombinant protein

Recommended Storage

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

Primary Application

In vitro research assays including SDS-PAGE, Western blot, and functional studies

Materials and Equipment Required

Before beginning the protocol, ensure the following materials and equipment are available:

• Recombinant Human C6orf163 Protein (Aladdin Scientific, SKU: rp376452)
• Sterile, nuclease-free water or the manufacturer-recommended reconstitution buffer
• Low-protein-binding microcentrifuge tubes (1.5 mL)
• Calibrated micropipettes and sterile tips
• Benchtop microcentrifuge
• Ice bucket or cooling block
• -20°C and/or -80°C freezer for long-term storage
• Appropriate downstream assay reagents (e.g., SDS-PAGE gel, Western blot apparatus)

All handling should be performed under aseptic conditions to prevent contamination and protease-mediated degradation.

Reconstitution Procedure

Follow these steps carefully to reconstitute Recombinant Human C6orf163 Protein without compromising its integrity:

1. Equilibrate the vial: Allow the lyophilized protein vial to equilibrate to room temperature for 10–15 minutes before opening. This minimizes moisture absorption from the environment.
2. Brief centrifugation: Briefly centrifuge the sealed vial (1,000–2,000 × g, 30 seconds) to consolidate the lyophilized material at the bottom of the tube.
3. Add reconstitution buffer: Slowly add the recommended volume of sterile reconstitution buffer (typically sterile deionized water or PBS, pH 7.4) along the inner wall of the vial. Do not pipette directly onto the lyophilized cake to avoid foaming and denaturation.
4. Gentle mixing: Gently swirl or pipette slowly to dissolve. Do not vortex, as shear forces can denature the protein.
5. Allow dissolution: Allow the vial to stand at room temperature for 5–10 minutes with occasional gentle mixing until the solution is homogeneous and clear.
6. Inspect the solution: Visually inspect for complete dissolution. The solution should be clear to slightly opalescent with no visible particulates.

Note: Refer to the specific product datasheet for the exact recommended reconstitution volume and buffer composition, as conditions may vary by lot.

Aliquoting and Storage

Proper aliquoting and storage are critical for preserving the biological activity of Recombinant Human C6orf163 Protein over time.

Aliquoting

• Immediately after reconstitution, divide the protein into single-use aliquots using low-protein-binding tubes to minimize adsorption losses.
• Aliquot volumes should correspond to the amount required for a single experiment to avoid multiple freeze-thaw cycles.
• Label each aliquot clearly with the protein name, concentration, date of reconstitution, and lot number.

Short-Term Storage

• For use within 1–2 weeks, store aliquots at 4°C. Adding a carrier protein such as 0.1% BSA (bovine serum albumin) may help stabilize the protein during short-term storage.

Long-Term Storage

• For long-term storage, snap-freeze aliquots in liquid nitrogen and store at -80°C. Storage at -20°C is acceptable for shorter periods.
• Repeated freeze-thaw cycles must be strictly avoided as they significantly reduce protein activity and increase aggregation.

Note: Specific storage conditions, recommended stabilizers, and shelf life should be confirmed against the product-specific datasheet provided with SKU rp376452.

Quality Control and Verification

Prior to use in critical downstream experiments, it is strongly recommended to verify the quality and integrity of the reconstituted Recombinant Human C6orf163 Protein.

• SDS-PAGE: Run a reducing SDS-PAGE gel to confirm the expected molecular weight band. Compare against the molecular weight indicated on the product datasheet.
• Protein concentration determination: Use a validated colorimetric assay (e.g., Bradford, BCA assay) to determine the actual protein concentration of the reconstituted solution, and compare against the stated nominal concentration.
• Western Blot: If an anti-C6orf163 antibody is available, confirm identity and purity by Western blot analysis.
• Visual inspection: Ensure the solution is free from visible aggregates, turbidity, or discoloration, which may indicate protein degradation.

Document all QC results and retain records alongside the lot number and reconstitution date for experimental traceability.

Downstream Applications and Usage Notes

Recombinant Human C6orf163 Protein is intended for research use only and is not suitable for therapeutic, diagnostic, or clinical applications. Typical in vitro research applications may include:

• Protein interaction studies: Use in pull-down assays or co-immunoprecipitation to investigate binding partners of C6orf163.
• Functional assays: Evaluate the biochemical activity of C6orf163 under defined assay conditions.
• Antibody validation: Serve as a positive control for anti-C6orf163 antibody validation by Western blot or ELISA.
• Structural studies: Use as substrate for structural characterization techniques.

General Usage Tips

• Keep protein on ice during all experimental manipulations to minimize degradation.
• Use the minimum concentration necessary to achieve experimental objectives to conserve material.
• Always include appropriate negative and positive controls in each assay.
• Consult the product datasheet for any application-specific recommendations provided by Aladdin Scientific.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Frequently asked questions

How should I reconstitute Recombinant Human C6orf163 Protein (rp376452)?

Allow the vial to equilibrate to room temperature, briefly centrifuge to collect the lyophilized material, then slowly add sterile reconstitution buffer (e.g., PBS pH 7.4) along the inner wall of the vial. Gently swirl to dissolve without vortexing. Refer to the product-specific datasheet for the exact recommended volume and buffer.

What are the recommended storage conditions for Recombinant Human C6orf163 Protein?

After reconstitution, aliquot the protein into single-use volumes using low-protein-binding tubes and store at -80°C for long-term storage or at 4°C for up to 1–2 weeks. Repeated freeze-thaw cycles should be strictly avoided to prevent loss of activity and aggregation.

How can I verify the quality of Recombinant Human C6orf163 Protein before use?

Run a reducing SDS-PAGE gel to confirm the expected molecular weight, and use a Bradford or BCA assay to determine the actual protein concentration. If an anti-C6orf163 antibody is available, Western blot analysis can further confirm identity and purity. Always inspect visually for aggregation or turbidity before use.

Can Recombinant Human C6orf163 Protein be used in clinical or diagnostic applications?

No. Recombinant Human C6orf163 Protein (rp376452) from Aladdin Scientific is strictly for research use only. It is not approved or validated for use in diagnostic, therapeutic, or clinical procedures.

Why should I avoid vortexing when reconstituting the protein?

Vortexing generates high shear forces that can denature or aggregate recombinant proteins, leading to loss of biological activity and potential formation of particulates. Gentle swirling or slow pipetting is recommended to dissolve the lyophilized material without compromising protein integrity.

References

1. Aladdin Scientific Product Datasheet: Recombinant Human C6orf163 Protein, SKU rp376452. Aladdin Scientific.
2. Deutscher MP (ed.). Guide to Protein Purification. Methods in Enzymology, Vol. 182. Academic Press, 1990.
3. Burgess RR, Deutscher MP (eds.). Guide to Protein Purification, 2nd ed. Methods in Enzymology, Vol. 463. Academic Press, 2009.

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