Recombinant Human CHKB Protein: FAQs & Technical Guide
Recombinant Human CHKB Protein: FAQs & Technical Guide
Recombinant Human CHKB Protein is a recombinant form of the human choline kinase beta enzyme, produced for research use to study phospholipid biosynthesis, kinase activity, and related metabolic and disease pathways.
- Product Name
- Recombinant Human CHKB Protein
- SKU
- rp375976
- Protein Function
- Catalyzes the phosphorylation of choline to phosphocholine, a key step in the CDP-choline (Kennedy) pathway for phosphatidylcholine biosynthesis
- Gene Name
- CHKB (also known as CHK, CHETK)
- Species
- Human (Homo sapiens)
- Application
- Biochemical assays, enzyme activity studies, drug discovery, phospholipid metabolism research
- Storage Recommendation
- Store at -80°C; avoid repeated freeze-thaw cycles (general recommendation for recombinant proteins; verify with datasheet)
What Is CHKB and Why Is It Important in Research?
CHKB (Choline Kinase Beta) is an enzyme encoded by the CHKB gene in humans. It catalyzes the first committed step in the CDP-choline (Kennedy) pathway: the ATP-dependent phosphorylation of choline to produce phosphocholine. This reaction is essential for the de novo biosynthesis of phosphatidylcholine (PC) and phosphatidylethanolamine (PE), the predominant phospholipids in eukaryotic cell membranes.
CHKB plays a critical role in maintaining membrane integrity and cellular phospholipid homeostasis. Mutations in the CHKB gene have been linked to congenital muscular dystrophy with mitochondrial structural abnormalities (megaconial type), underscoring its physiological importance. Additionally, choline kinase activity has attracted attention in oncology research, as altered phospholipid metabolism is a hallmark of many cancer cell types.
Recombinant Human CHKB Protein enables researchers to study kinase kinetics, screen inhibitors, and investigate the molecular basis of CHKB-associated diseases in a controlled in vitro setting.
Product Specifications and Quality
Aladdin Scientific's Recombinant Human CHKB Protein (SKU: rp375976) is designed for use in biochemical and cell-free research applications. While full specification details should always be confirmed in the current product datasheet, the following general guidance applies to recombinant kinase proteins of this type:
- Expression System: Typically produced in a prokaryotic (e.g., E. coli) or eukaryotic (e.g., insect or HEK293) expression system. Please consult the product datasheet for confirmed details.
- Purity: Recombinant proteins from Aladdin are generally supplied at >85–95% purity as assessed by SDS-PAGE. Confirm exact purity on the product certificate of analysis (CoA).
- Tag: May include an affinity purification tag (e.g., His-tag) to facilitate purification; confirm tag identity and position on the datasheet.
- Formulation: Typically supplied lyophilized or in a stabilizing buffer. Check the product datasheet for formulation details.
- Activity: Functional activity (e.g., kinase activity toward choline substrate) may be validated; refer to the CoA for lot-specific data.
Note: All specific numerical values (molecular weight, exact purity percentage, endotoxin level, activity units) must be verified against the product datasheet and CoA for lot rp375976.
Storage, Handling, and Reconstitution
Proper storage and handling of Recombinant Human CHKB Protein is critical to maintaining its biological activity and experimental reproducibility. The following guidelines reflect best practices for recombinant proteins; always defer to the specific instructions provided in the product datasheet.
Storage
- Store the protein at -80°C for long-term stability.
- For short-term use (up to 1–2 weeks), storage at -20°C may be acceptable depending on formulation.
- Avoid repeated freeze-thaw cycles, as these can lead to protein aggregation, loss of activity, and degradation. Aliquot upon first use.
Reconstitution (if lyophilized)
- Allow the vial to equilibrate to room temperature before opening to minimize condensation.
- Reconstitute with the volume of sterile water or buffer recommended in the datasheet to achieve the desired concentration.
- Gently pipette or swirl to dissolve; do not vortex vigorously.
- Allow 10–15 minutes for complete reconstitution, then centrifuge briefly before use.
Handling
- Keep on ice during experiments.
- Use low-protein-binding tubes and pipette tips to minimize adsorption losses.
- Addition of a carrier protein (e.g., 0.1% BSA) to dilution buffers may help stabilize the protein during extended assays.
Research Applications of Recombinant Human CHKB Protein
Recombinant Human CHKB Protein is a versatile reagent suitable for multiple research contexts:
- Enzyme Kinetics Studies: Determine kinetic parameters (Km, Vmax, kcat) for choline phosphorylation using standard radioactive or fluorescence-based kinase assays.
- Inhibitor Screening: Use as a target enzyme in high-throughput or focused screens to identify small-molecule inhibitors of choline kinase activity, relevant to cancer metabolism and muscular dystrophy research.
- Structural and Biophysical Studies: Employ in crystallography, cryo-EM, or biophysical assays (e.g., SPR, ITC) to characterize CHKB structure and substrate interactions.
- Antibody Validation: Serve as a positive control antigen for validating anti-CHKB antibodies in Western blot or ELISA formats.
- Disease Mechanism Research: Investigate molecular mechanisms underlying CHKB-associated congenital muscular dystrophy (megaconial type) and other phospholipid metabolism disorders.
- Drug Discovery: Support target engagement assays and structure-activity relationship (SAR) studies for therapeutic programs targeting the Kennedy pathway.
Troubleshooting Common Issues
If you experience unexpected results with Recombinant Human CHKB Protein, consider the following troubleshooting steps:
| Issue | Possible Cause | Suggested Action |
|---|---|---|
| No or low enzyme activity | Protein degradation due to improper storage or freeze-thaw | Use freshly aliquoted protein; verify storage conditions; check CoA activity data |
| Protein precipitation or aggregation | Incorrect reconstitution or incompatible buffer | Follow datasheet reconstitution protocol; use recommended buffer and gentle mixing |
| Unexpected band size on SDS-PAGE | Tag contribution to molecular weight; post-translational modifications; glycosylation if eukaryotic expression | Compare with theoretical MW including tag; consult datasheet |
| High background in kinase assay | ATP contamination or non-specific phosphorylation | Include no-enzyme control; verify substrate and ATP concentrations |
| Inconsistent results between lots | Lot-to-lot variability | Request CoA for each lot; run internal reference standard in parallel |
For unresolved issues, contact Aladdin Scientific technical support with your lot number, CoA data, and experimental protocol details.
Frequently asked questions
What is the biological function of the CHKB protein?
CHKB (Choline Kinase Beta) catalyzes the ATP-dependent phosphorylation of choline to phosphocholine, the first committed step of the CDP-choline (Kennedy) pathway for phosphatidylcholine biosynthesis. This pathway is essential for de novo membrane phospholipid production in eukaryotic cells.
What expression system is used to produce Recombinant Human CHKB Protein (rp375976)?
The specific expression system for rp375976 should be confirmed in the product datasheet available from Aladdin Scientific. Recombinant CHKB proteins are commonly expressed in E. coli, insect cells (baculovirus), or mammalian cell systems depending on the supplier's production process and intended application.
How should I store Recombinant Human CHKB Protein to preserve its activity?
For long-term storage, keep the protein at -80°C. Aliquot the protein upon first use to avoid repeated freeze-thaw cycles, which can cause aggregation and loss of enzymatic activity. If the protein is lyophilized, store it desiccated at the recommended temperature until reconstitution.
What research applications is Recombinant Human CHKB Protein suitable for?
Recombinant Human CHKB Protein is suitable for enzyme kinetics studies, inhibitor screening, structural and biophysical characterization, antibody validation (as a positive control antigen), and investigation of CHKB-related disease mechanisms such as megaconial congenital muscular dystrophy. It is also used in drug discovery programs targeting phospholipid metabolism.
Is Recombinant Human CHKB Protein suitable for use in cell-based assays?
Recombinant Human CHKB Protein is primarily intended for cell-free biochemical assays such as enzyme activity measurements, inhibitor screens, and biophysical studies. Suitability for cell-based applications depends on the delivery method and experimental design; please consult the product datasheet or contact Aladdin Scientific technical support for guidance specific to your application.
What diseases are associated with CHKB gene mutations?
Mutations in the CHKB gene are associated with megaconial congenital muscular dystrophy (also referred to as CHKB-related muscular dystrophy), a rare autosomal recessive disorder characterized by muscle weakness and mitochondrial structural abnormalities (enlarged mitochondria in muscle fibers). Altered choline kinase activity has also been studied in the context of cancer metabolism.
References
- Aladdin Scientific. Recombinant Human CHKB Protein (rp375976). Product datasheet. Aladdin Scientific.
- Aoyama C, Liao H, Ishidate K. Structure and function of choline kinase isoforms in mammalian cells. Prog Lipid Res. 2004;43(3):266-281.
- Mak BC, et al. Mutations in CHKB cause megaconial congenital muscular dystrophy. Nature Genetics. 2011.
- Glunde K, Bhujwalla ZM, Bhujwalla S. Choline metabolism in malignant transformation. Nat Rev Cancer. 2011;11(12):835-848.
Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.
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