Recombinant Human GLS2 Protein: Research Guide & Applications

Recombinant Human GLS2 Protein is a liver-type glutaminase enzyme critical for glutamine catabolism, tumor suppression, and neurological function, offering researchers a well-characterized tool for in vitro biochemical and functional studies.

Protein Name

Recombinant Human GLS2 (Glutaminase 2, Liver-type)

SKU

rp374603

Species

Human (Homo sapiens)

Protein Function

Catalyzes the hydrolysis of glutamine to glutamate and ammonia; acts as a tumor suppressor regulated by p53

Primary Research Areas

Glutamine metabolism, cancer biology, neuroscience, metabolic disease

Recommended Storage

Store at -80°C; avoid repeated freeze-thaw cycles

Typical Applications

Enzyme activity assays, Western blot, ELISA, protein interaction studies

Introduction to GLS2 and Its Biological Significance

Glutaminase 2 (GLS2), also known as liver-type glutaminase, is a mitochondrial enzyme encoded by the GLS2 gene in humans. It catalyzes the hydrolytic conversion of glutamine to glutamate and ammonia, a critical step in glutamine catabolism that feeds into the tricarboxylic acid (TCA) cycle and supports cellular bioenergetics, nucleotide synthesis, and redox homeostasis.

Unlike the kidney-type isoform GLS1, GLS2 is predominantly expressed in the liver and brain. Importantly, GLS2 has been identified as a transcriptional target of the tumor suppressor p53, linking glutamine metabolism directly to oncogenic regulation. Loss of GLS2 expression is associated with hepatocellular carcinoma and other malignancies, underscoring its role as a metabolic tumor suppressor. In neurological contexts, GLS2 contributes to glutamatergic neurotransmitter pools and overall nitrogen homeostasis.

Recombinant Human GLS2 Protein (SKU: rp374603) provides researchers with a well-characterized, ready-to-use enzyme for exploring these biological pathways in controlled in vitro settings.

Reconstitution and Handling Protocol

Proper handling of Recombinant Human GLS2 Protein is essential to preserve enzymatic activity and experimental reproducibility. Follow these guidelines carefully:

• Centrifugation before opening: Briefly centrifuge the vial at low speed before opening to collect any lyophilized material from the walls and cap.
• Reconstitution buffer: Reconstitute the lyophilized protein in sterile deionized water or an appropriate buffer such as phosphate-buffered saline (PBS, pH 7.4). The specific reconstitution volume and recommended buffer should be confirmed from the product datasheet (absea #PP-8387).
• Gentle mixing: Gently pipette or swirl the solution to dissolve the protein. Avoid vigorous vortexing, which may denature the protein.
• Protein concentration: Determine the working concentration using a validated protein quantification method (e.g., BCA or Bradford assay) prior to use in assays.
• Aliquoting: After reconstitution, divide the solution into single-use aliquots to minimize freeze-thaw cycles, which can reduce enzymatic activity and protein integrity.

Note: Specific reconstitution volumes, buffer compositions, and carrier protein requirements should be verified against the official product datasheet prior to use.

Storage, Stability, and Quality Control

Maintaining the stability of Recombinant Human GLS2 Protein requires adherence to recommended storage conditions. Key guidelines include:

• Long-term storage: Store at -80°C in single-use aliquots. Under these conditions, the protein is expected to remain stable for up to 12 months from the date of receipt, though the exact shelf life should be confirmed on the product datasheet.
• Short-term storage: If the protein will be used within a few days, it may be stored at 4°C. Extended storage at 4°C is not recommended.
• Avoid freeze-thaw cycles: Repeated freezing and thawing significantly reduces protein activity and can cause aggregation. Always pre-aliquot upon first reconstitution.
• Carrier proteins: Addition of a stabilizing carrier protein (e.g., 0.1% BSA) to the reconstitution buffer may improve stability during storage, depending on the downstream application.

Aladdin Scientific recommends verifying protein activity and purity upon receipt using SDS-PAGE or activity-based assays to establish a baseline before commencing experiments. All quality control data should be referenced from the official Certificate of Analysis (CoA) shipped with the product.

Research Applications of Recombinant Human GLS2 Protein

Recombinant Human GLS2 Protein is a versatile reagent suited to a broad range of experimental applications in biochemistry, cell biology, and translational research:

• Enzyme activity assays: Quantify glutaminase activity by measuring the conversion of glutamine to glutamate using colorimetric or fluorometric assay kits. GLS2 protein serves as a positive control or study substrate for inhibitor screening campaigns.
• Inhibitor and drug discovery: Use purified GLS2 protein in high-throughput screening (HTS) or structure-activity relationship (SAR) studies to identify and characterize small-molecule inhibitors of glutamine metabolism, relevant to anticancer drug development.
• Western blot and ELISA: Recombinant GLS2 can serve as a standard or positive control in immunoassays to validate antibody specificity and assay sensitivity.
• Protein-protein interaction studies: Employ GLS2 in pull-down assays, co-immunoprecipitation (co-IP), or surface plasmon resonance (SPR) experiments to map interaction networks involving glutamine metabolism regulators.
• Structural biology: Use recombinant GLS2 as a substrate for crystallography or cryo-EM studies aimed at elucidating enzyme structure and catalytic mechanism.
• p53 pathway research: Investigate the regulatory axis between p53 and GLS2 in the context of cancer metabolism and oxidative stress response.

Frequently Encountered Troubleshooting Considerations

To ensure optimal performance of Recombinant Human GLS2 Protein in your experiments, consider the following troubleshooting guidance:

• Low or no enzymatic activity: Verify that the protein was stored and reconstituted correctly. Check that the assay buffer pH and temperature are within the optimal range for GLS2 activity. Confirm that the protein has not undergone excessive freeze-thaw cycles.
• Protein aggregation or precipitation: If visible aggregates are observed, gently centrifuge the reconstituted solution at low speed and use the supernatant. Consider adding a carrier protein such as BSA to the storage buffer.
• Inconsistent assay results: Ensure thorough mixing of the reconstituted protein prior to aliquoting and use. Standardize pipetting technique and confirm reagent lot consistency.
• Weak signal in immunoassays: Confirm antibody compatibility with the recombinant protein's expression tag or epitope. Use the recombinant protein as a positive control to validate antibody performance before testing biological samples.

For product-specific technical support, consult the official product datasheet (absea #PP-8387) or contact Aladdin Scientific technical support directly.

Frequently asked questions

What is GLS2 and why is it important in cancer research?

GLS2 (Glutaminase 2) is a liver-type mitochondrial enzyme that converts glutamine to glutamate, fueling the TCA cycle and cellular energy production. It is a direct transcriptional target of the tumor suppressor p53, and its expression is frequently downregulated in hepatocellular carcinoma and other cancers, making it a key focus of cancer metabolism and drug discovery research.

How should I reconstitute Recombinant Human GLS2 Protein for my experiment?

Reconstitute the lyophilized GLS2 protein in sterile deionized water or an appropriate buffer such as PBS (pH 7.4). Gently pipette to mix and avoid vigorous vortexing. Always refer to the product datasheet (absea #PP-8387) for the recommended reconstitution volume and any specific buffer requirements for your application.

What are the recommended storage conditions for Recombinant Human GLS2 Protein?

Store Recombinant Human GLS2 Protein at -80°C in single-use aliquots to maximize stability and prevent loss of enzymatic activity from repeated freeze-thaw cycles. Short-term storage at 4°C is acceptable if the protein will be used within a few days. Confirm the exact shelf life from the product Certificate of Analysis.

Can Recombinant Human GLS2 Protein be used as a positive control in Western blot or ELISA?

Yes, Recombinant Human GLS2 Protein (SKU: rp374603) is well-suited for use as a positive control or standard in immunoassays including Western blot and ELISA. It enables validation of antibody specificity and helps establish assay sensitivity before testing complex biological samples.

Is Recombinant Human GLS2 Protein suitable for inhibitor screening assays?

Yes, purified Recombinant Human GLS2 Protein is appropriate for use in enzyme inhibitor screening, including high-throughput screening (HTS) and structure-activity relationship (SAR) studies targeting glutamine metabolism pathways relevant to anticancer drug discovery. Ensure assay buffer conditions are optimized for GLS2 enzymatic activity prior to screening.

References

1. Matés JM, et al. Glutaminase isoenzymes as key regulators in metabolic and oxidative stress against cancer. Curr Mol Med. 2013.
2. Hu W, et al. Glutaminase 2, a novel p53 target gene regulating energy metabolism and antioxidant function. Proc Natl Acad Sci USA. 2010.
3. Suzuki S, et al. Phosphate-activated glutaminase (GLS2), a p53-inducible regulator of glutamine metabolism and reactive oxygen species. Proc Natl Acad Sci USA. 2010.
4. Aladdin Scientific. Recombinant Human GLS2 Protein (rp374603) Product Datasheet. absea #PP-8387.

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