Recombinant Human CHCT1 Protein: Production & Usage Protocol

Recombinant Human CHCT1 Protein is a research-grade recombinant protein intended for use in biochemical, binding, and cell-based assays requiring high-purity human CHCT1.

Product Name

Recombinant Human CHCT1 Protein

SKU

rp370207

Species

Human (Homo sapiens)

Protein Type

Recombinant protein

Intended Use

Research use only (RUO)

Storage Recommendation

Avoid repeated freeze-thaw cycles; store as recommended on product datasheet

Overview of Recombinant Human CHCT1 Protein

Recombinant Human CHCT1 Protein (SKU: rp370207) is produced for research applications requiring a well-characterized, human-derived recombinant protein. CHCT1 (also referred to in some literature contexts as a protein involved in cellular regulatory processes) is expressed using recombinant technology to ensure batch-to-batch consistency and high purity suitable for demanding biochemical and cell-based experiments.

This product is intended for research use only and is not approved for diagnostic or therapeutic applications. Researchers should consult the product datasheet for the most current information on purity, formulation, and recommended assay conditions.

Materials and Equipment Required

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

• Recombinant Human CHCT1 Protein (rp370207, Aladdin Scientific)
• Sterile, nuclease-free water or appropriate reconstitution buffer (as specified on the product datasheet)
• Microcentrifuge tubes (low-binding, 1.5 mL recommended)
• Calibrated micropipettes and sterile tips
• Microcentrifuge
• Ice or cooling block for sample handling
• Appropriate assay buffers and reagents for downstream applications
• Protein quantification reagents (e.g., BCA or Bradford assay kit)

All equipment should be calibrated and cleaned prior to use to avoid cross-contamination and ensure reproducibility.

Reconstitution and Handling Protocol

Follow the steps below for proper reconstitution and handling of Recombinant Human CHCT1 Protein:

1. Thawing: Remove the vial from storage and allow it to equilibrate at room temperature for no more than 5 minutes, or thaw briefly on ice. Do not thaw at 37 °C to prevent protein degradation.
2. Reconstitution: Briefly centrifuge the vial to collect any lyophilized material at the bottom. Add the volume of sterile reconstitution buffer specified on the product datasheet. Gently pipette up and down to mix; do not vortex vigorously, as this may cause protein denaturation.
3. Equilibration: Allow the reconstituted solution to sit on ice for 5–10 minutes to ensure complete dissolution.
4. Protein Quantification: Determine protein concentration using a validated assay (e.g., BCA or Bradford) according to the manufacturer's instructions, and compare against the expected concentration stated on the datasheet.
5. Aliquoting: Prepare single-use aliquots to avoid repeated freeze-thaw cycles, which can reduce protein activity and integrity.
6. Storage: Store aliquots according to the conditions specified on the product datasheet. Generally, short-term storage at 4 °C (up to 1 week) and long-term storage at −20 °C or −80 °C are recommended. Avoid repeated freeze-thaw cycles.

Note: Always refer to the product-specific datasheet provided with each lot for reconstitution volume, buffer composition, and storage conditions, as these may vary by formulation.

Quality Control and Verification

Quality control (QC) is essential to confirm that the recombinant protein is suitable for intended experimental applications. The following QC steps are recommended:

• SDS-PAGE: Resolve the reconstituted protein by SDS-PAGE under reducing and non-reducing conditions to verify molecular weight and purity. Compare the observed band with the expected molecular weight stated on the datasheet.
• Western Blot: Confirm protein identity using a validated anti-CHCT1 antibody. A single specific band at the expected molecular weight indicates correct protein identity.
• Protein Concentration: Compare the measured concentration against the lot-specific certificate of analysis (CoA) value.
• Functional Testing: Where applicable, perform a functional assay (e.g., binding assay, enzyme activity assay) to confirm biological activity prior to use in critical experiments.

Document all QC results and retain the CoA and datasheet for each lot used in experiments to support data reproducibility and traceability.

Troubleshooting and Best Practices

If unexpected results are encountered, consult the following troubleshooting guide:

• Low protein recovery after reconstitution: Ensure the full recommended volume of reconstitution buffer is added. Allow additional time (up to 15 minutes on ice) for complete dissolution. Check that the vial was centrifuged prior to opening.
• Multiple bands on SDS-PAGE: This may indicate protein aggregation or degradation. Confirm that the protein was not exposed to elevated temperatures or repeated freeze-thaw cycles. Re-run under both reducing and non-reducing conditions.
• Loss of biological activity: Verify storage conditions and total number of freeze-thaw cycles. Prepare fresh aliquots if activity loss is suspected.
• High background in downstream assays: Ensure buffers are compatible with the protein formulation. Optimize blocking conditions and antibody concentrations.

For additional technical support, contact Aladdin Scientific's technical support team with the lot number and datasheet information.

Frequently asked questions

How should Recombinant Human CHCT1 Protein (rp370207) be stored after reconstitution?

After reconstitution, prepare single-use aliquots and store them according to the product datasheet. Generally, short-term storage at 4 °C (up to one week) and long-term storage at −20 °C or −80 °C are recommended. Avoid repeated freeze-thaw cycles to preserve protein integrity and activity.

What reconstitution buffer should be used for Recombinant Human CHCT1 Protein?

The recommended reconstitution buffer and volume are specified on the product datasheet and certificate of analysis provided with each lot. Always refer to these lot-specific documents before reconstituting the protein, as formulations may vary between lots.

How can I verify the identity and purity of the reconstituted CHCT1 protein?

Identity and purity can be confirmed by SDS-PAGE to check molecular weight and band purity, followed by Western blot using a validated anti-CHCT1 antibody to confirm protein identity. Protein concentration should be measured using a validated assay such as BCA or Bradford and compared against the certificate of analysis.

Is Recombinant Human CHCT1 Protein suitable for clinical or diagnostic use?

No. Recombinant Human CHCT1 Protein (rp370207) is intended for research use only (RUO) and is not approved or validated for diagnostic, therapeutic, or clinical applications.

What should I do if I observe multiple bands or reduced activity after reconstitution?

Multiple bands may indicate protein aggregation or degradation due to improper storage or excessive freeze-thaw cycles. Verify that storage conditions were followed correctly and that the protein was not exposed to elevated temperatures. Prepare fresh aliquots and contact Aladdin Scientific technical support with your lot number if issues persist.

References

1. Product Datasheet: Recombinant Human CHCT1 Protein, Aladdin Scientific, SKU rp370207. Refer to lot-specific certificate of analysis for detailed specifications.
2. Scopes, R.K. Protein Purification: Principles and Practice. 3rd ed. Springer, New York, 1994. (General reference for recombinant protein handling and QC.)

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