Recombinant Human CHCHD6 Protein: Properties & Lab Guide

Recombinant Human CHCHD6 Protein is a research-grade recombinant protein targeting the human CHCHD6 gene product, a mitochondria-associated coiled-coil-helix-coiled-coil-helix domain-containing protein implicated in mitochondrial inner membrane organization and cristae morphology maintenance.

Product Name

Recombinant Human CHCHD6 Protein

SKU

rp375211

Protein Family

CHCHD (Coiled-Coil-Helix-Coiled-Coil-Helix Domain) family

Biological Origin

Human (Homo sapiens)

Primary Research Application

Mitochondrial biology, protein-protein interaction studies, functional proteomics

Storage Recommendation

Aliquot and store at -80 °C; avoid repeated freeze-thaw cycles

Supplier

Aladdin Scientific

What Is CHCHD6? Biological Background

CHCHD6 (Coiled-Coil-Helix-Coiled-Coil-Helix Domain Containing 6) is a nuclear-encoded mitochondrial protein belonging to the CHCHD protein superfamily. Proteins in this family are characterized by conserved CHCH structural motifs that coordinate zinc ions or form disulfide bonds important for protein folding and stability within the mitochondrial intermembrane space or inner membrane.

CHCHD6 has been reported to associate with the mitochondrial inner membrane contact site and cristae organizing system (MINOS/MICOS complex), which plays a critical role in maintaining mitochondrial cristae architecture, membrane dynamics, and overall organelle integrity. Disruption of MICOS complex components, including CHCHD6, can lead to fragmented or aberrant cristae morphology, impaired oxidative phosphorylation, and altered mitochondrial signaling.

Research into CHCHD6 contributes to a broader understanding of mitochondrial quality control, bioenergetics, and the pathological mechanisms underlying mitochondria-associated diseases, including neurodegeneration and metabolic disorders.

Product Specifications & Quality Attributes

Aladdin Scientific's Recombinant Human CHCHD6 Protein (SKU: rp375211) is produced using recombinant expression systems to deliver a research-grade protein suitable for a range of in vitro biochemical and cell biology assays. Key quality attributes researchers should verify upon receipt include:

• Purity: Recombinant proteins are typically assessed by SDS-PAGE and/or HPLC to confirm purity. Researchers should consult the Certificate of Analysis (CoA) provided with each lot for confirmed purity percentage.
• Formulation: The protein is supplied in a defined buffer formulation designed to maintain solubility and stability. Exact buffer composition is detailed in the product datasheet; researchers should verify compatibility with their assay conditions.
• Biological Activity: Functional validation may include binding assays or activity assays relevant to CHCHD6's known interactions. Consult the product datasheet for lot-specific activity data.
• Endotoxin Level: For cell-based assays, endotoxin content is a critical quality parameter. Refer to the CoA for endotoxin test results.

All lot-specific data should be verified against the CoA shipped with the product. Contact Aladdin Scientific technical support for additional characterization data if required.

Handling, Reconstitution & Storage Protocol

Proper handling of Recombinant Human CHCHD6 Protein is essential to preserve activity and ensure reproducible experimental results. Follow the guidelines below:

1. Upon Receipt: Inspect the shipment immediately. If the protein is supplied lyophilized, confirm the vial is intact and the cake is present. If supplied in liquid form, confirm the vial is sealed and has remained sufficiently cold during transit.
2. Reconstitution (Lyophilized Form): Briefly centrifuge the vial before opening to consolidate the powder. Reconstitute in the recommended volume of sterile, deionized water or the buffer specified in the datasheet. Allow the protein to dissolve gently at room temperature or on ice; do not vortex vigorously, as this may cause aggregation or denaturation.
3. Aliquoting: Prepare single-use aliquots immediately after reconstitution to minimize freeze-thaw cycles. Label each aliquot with the date of reconstitution and lot number.
4. Storage: Store aliquots at -80 °C for long-term storage. For short-term use (up to 1 week), aliquots may be stored at -20 °C. Avoid repeated freeze-thaw cycles, which degrade protein integrity and reduce activity.
5. Working Concentration: Determine the optimal working concentration empirically for each assay type. Refer to published literature on CHCHD6 or MICOS complex studies as a starting reference.

Note: Specific reconstitution volumes, buffer compositions, and storage conditions may vary by lot. Always refer to the product-specific datasheet and CoA for authoritative guidance.

Research Applications

Recombinant Human CHCHD6 Protein supports a variety of research applications in the fields of mitochondrial biology and cell biology. Representative applications include:

• Protein-Protein Interaction Studies: CHCHD6 is a component of the MICOS/MINOS complex. The recombinant protein can be used in pull-down assays, co-immunoprecipitation controls, or surface plasmon resonance (SPR) experiments to map interaction partners within the complex or identify novel binding proteins.
• Antibody Validation: Purified recombinant CHCHD6 protein serves as a positive control antigen for validating anti-CHCHD6 antibodies used in Western blot, ELISA, or immunofluorescence assays.
• Biochemical Assays: The protein can be used to study CHCHD6 enzymatic or structural properties in defined in vitro systems, contributing to mechanistic understanding of MICOS complex assembly and function.
• Assay Development & Screening: Recombinant CHCHD6 can serve as a target protein in drug discovery screening campaigns aimed at identifying small molecules that modulate MICOS complex integrity or mitochondrial dynamics.
• Structural Biology: High-purity recombinant protein preparations are suitable as starting material for crystallography or cryo-electron microscopy studies aimed at resolving CHCHD6 three-dimensional structure.

Troubleshooting & Technical Tips

Researchers working with Recombinant Human CHCHD6 Protein may encounter challenges common to recombinant protein work. The following troubleshooting guidance is provided:

• Protein Aggregation: If aggregation is observed upon reconstitution, try dissolving the protein at a lower concentration or in a buffer containing a mild detergent (e.g., 0.01–0.05% Tween-20) or a low concentration of arginine (50–100 mM) to improve solubility. Keep the protein on ice throughout.
• Loss of Activity After Freeze-Thaw: Activity loss is often caused by repeated freeze-thaw cycles. Ensure single-use aliquots are prepared at the time of reconstitution. Adding carrier protein (e.g., 0.1% BSA) to storage buffer may help stabilize dilute protein preparations.
• Unexpected Band Size on SDS-PAGE: CHCHD6 may run at an apparent molecular weight slightly different from its calculated sequence-based molecular weight due to post-translational modifications or the presence of affinity tags. Consult the datasheet for expected band size and tag information.
• Low Signal in Binding Assays: Optimize protein concentration, incubation time, and buffer conditions. Ensure the recombinant protein is not denatured prior to the assay; native conformation is critical for interaction studies.

For additional technical support, contact Aladdin Scientific's technical support team with your lot number and CoA details.

Frequently asked questions

What expression system is used to produce Recombinant Human CHCHD6 Protein (rp375211)?

The specific expression system (e.g., E. coli, baculovirus/insect cells, mammalian cells, or wheat germ) for rp375211 should be confirmed in the product datasheet or Certificate of Analysis provided by Aladdin Scientific. Different expression systems may affect post-translational modifications and protein folding; please contact Aladdin Scientific technical support for lot-specific details.

How should I validate that Recombinant Human CHCHD6 Protein is functional in my assay?

Functional validation strategies depend on your assay format. For protein interaction studies, you can test binding to known MICOS complex partners. For antibody validation, run the recombinant protein alongside lysate controls on Western blot. Always include a positive control and refer to published literature on CHCHD6 biology to benchmark your results. Lot-specific activity data from Aladdin Scientific's CoA provides a starting reference point.

Can Recombinant Human CHCHD6 Protein be used directly in cell-based assays?

Yes, recombinant proteins can be used in cell-based assays, but endotoxin levels must be carefully controlled to avoid confounding inflammatory or cytotoxic effects. Always verify the endotoxin content from the CoA before use in cell culture. Additionally, confirm that the protein formulation buffer is compatible with your cell culture media and that protein concentrations used are within a non-toxic range for your cell type.

What is the role of CHCHD6 in the MICOS complex?

CHCHD6 is reported to be a component of the Mitochondrial Inner membrane Organizing System (MICOS) complex, which is essential for maintaining mitochondrial cristae architecture and inner membrane contact sites. Its CHCH domain is thought to contribute to complex stability and membrane tethering functions. The precise molecular mechanism by which CHCHD6 contributes to MICOS assembly continues to be an area of active investigation.

Where can I find the Certificate of Analysis (CoA) for my lot of rp375211?

The Certificate of Analysis for your specific lot of Recombinant Human CHCHD6 Protein (rp375211) can be obtained from Aladdin Scientific's website by entering your lot number, or by contacting Aladdin Scientific customer support directly. The CoA contains lot-specific purity, activity, endotoxin, and formulation data essential for experimental planning.

References

1. van der Laan M, Horvath SE, Pfanner N. Mitochondrial contact site and cristae organizing system. Curr Opin Cell Biol. 2016;41:33-42. doi:10.1016/j.ceb.2016.03.013
2. Pfanner N, van der Laan M, Amati P, et al. Uniform nomenclature for the mitochondrial contact site and cristae organizing system. J Cell Biol. 2014;204(7):1083-1086. doi:10.1083/jcb.201401006
3. Hoppins S, Collins SR, Bhatt A, et al. A mitochondrial-focused genetic interaction map reveals a scaffold-like complex required for inner membrane organization in mitochondria. J Cell Biol. 2011;195(2):323-340. doi:10.1083/jcb.201107053

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