Recombinant Human GLP2R Protein: Handling & Application Protocol

Recombinant Human GLP2R Protein is a purified recombinant form of the human glucagon-like peptide-2 receptor, designed to support GPCR binding assays, functional studies, and metabolic disease research with high batch-to-batch consistency.

Product Name

Recombinant Human GLP2R Protein

SKU

rp370014

Target

GLP2R (Glucagon-Like Peptide 2 Receptor)

Species

Human

Application

Binding assays, functional studies, GPCR research, metabolic disease studies

Storage

Store lyophilized protein at -20°C or -80°C; avoid repeated freeze-thaw cycles

Background and Biological Significance of GLP2R

The glucagon-like peptide-2 receptor (GLP2R) is a class B G protein-coupled receptor (GPCR) that is primarily expressed in intestinal enteroendocrine cells, enteric neurons, and subepithelial myofibroblasts. GLP2R is activated by its endogenous ligand, glucagon-like peptide-2 (GLP-2), a 33-amino acid peptide secreted from intestinal L-cells in response to nutrient intake.

Upon GLP-2 binding, GLP2R signals predominantly through the Gs pathway, stimulating adenylyl cyclase activity and increasing intracellular cAMP levels. This signaling cascade promotes intestinal epithelial proliferation, enhances gut barrier function, reduces epithelial apoptosis, and increases nutrient absorption. GLP2R is recognized as an important therapeutic target for short bowel syndrome, inflammatory bowel disease, and other intestinal disorders. Teduglutide, a GLP-2 analogue, has been approved for clinical use in short bowel syndrome, underscoring the translational relevance of GLP2R research.

Recombinant Human GLP2R Protein (rp370014) enables in vitro biochemical and cell-free studies of receptor structure, ligand binding, and downstream signaling, providing researchers with a valuable tool for drug discovery and mechanistic investigations.

Reconstitution and Storage Instructions

Reconstitution:

• Briefly centrifuge the vial at low speed (e.g., 1,000–2,000 × g for 30 seconds) before opening to collect lyophilized material at the bottom of the tube.
• Reconstitute the lyophilized protein in sterile, ultra-pure water or an appropriate buffer (e.g., PBS, pH 7.4) to a working stock concentration. Refer to the Certificate of Analysis (CoA) supplied with the product for the recommended reconstitution volume.
• Allow the protein to dissolve gently by pipetting up and down or by gentle swirling. Do NOT vortex, as vigorous agitation may cause protein denaturation or aggregation.
• If the protein will not be used immediately, add a carrier protein (e.g., 0.1% BSA) to stabilize the reconstituted solution.

Storage:

• Store the lyophilized protein at -20°C or -80°C upon receipt.
• After reconstitution, aliquot into single-use volumes to avoid repeated freeze-thaw cycles, which can lead to loss of activity and protein aggregation.
• Reconstituted protein aliquots should be stored at -80°C for long-term storage or at -20°C for short-term use (up to 1 month).
• Avoid exposure to repeated freeze-thaw cycles (maximum 3 cycles recommended).

Note: Specific concentration, purity, and reconstitution recommendations will be provided on the product CoA. Always refer to the CoA for lot-specific guidance.

Quality Verification and Handling Precautions

Quality Verification:

Before use, verify the identity and integrity of the Recombinant Human GLP2R Protein by the following methods:

• SDS-PAGE: Resolve a small aliquot (1–5 µg) under reducing or non-reducing conditions to confirm the apparent molecular weight and purity. Consult the CoA for the expected molecular weight band.
• Western Blot: Use a validated anti-GLP2R antibody to confirm protein identity.
• Protein Concentration Determination: Measure protein concentration using BCA or Bradford assay prior to experiments. Note that absorbance-based assays may be affected by buffer components; select the appropriate assay accordingly.

Handling Precautions:

• Always work on ice when handling reconstituted protein to minimize activity loss.
• Use low-binding microcentrifuge tubes and pipette tips to minimize protein adsorption to plastic surfaces.
• Wear appropriate PPE (gloves, lab coat, eye protection) when handling recombinant proteins.
• This product is intended for research use only (RUO) and is not suitable for therapeutic or diagnostic use in humans.

Experimental Applications

Recombinant Human GLP2R Protein (rp370014) is suitable for a range of in vitro research applications, including but not limited to:

• Ligand Binding Assays: Used to study the interaction between GLP-2 or GLP-2 analogues (e.g., teduglutide) and the GLP2R. Suitable for competitive binding assays and saturation binding studies to determine affinity constants (Kd) and binding kinetics.
• ELISA and Immunoassay Development: GLP2R protein can serve as a coating antigen or capture reagent for the development and validation of immunoassays detecting GLP2R or anti-GLP2R antibodies.
• Antibody Screening and Validation: Use as a positive control antigen or immunogen for generating and validating anti-GLP2R antibodies in hybridoma screening or phage display platforms.
• Drug Discovery and Screening: Incorporated into high-throughput binding or competition assays for identifying small-molecule or biologic modulators of GLP2R activity.
• Structural and Biochemical Studies: Supports biophysical characterization of the receptor protein, including thermal stability assays (DSF/TSA), SPR-based interaction studies, and pull-down assays.

Optimize protein concentrations and assay conditions for each specific application. Refer to relevant literature and the product CoA for recommended starting conditions.

Troubleshooting Common Issues

The following table summarizes common issues encountered when working with Recombinant Human GLP2R Protein and suggested remedies:

• No signal or weak signal in binding assay: Verify protein integrity by SDS-PAGE; confirm reconstitution was performed correctly; check that the protein was not subjected to excessive freeze-thaw cycles; ensure assay conditions (pH, salt concentration, temperature) are optimized for GLP2R.
• Protein precipitation or aggregation: Centrifuge reconstituted protein at 10,000–14,000 × g for 10 minutes at 4°C to remove aggregates before use; consider adding a non-ionic detergent (e.g., 0.01% Tween-20) or a stabilizing agent (e.g., glycerol, BSA) to the buffer; reduce protein concentration in the working solution.
• Unexpected band size on SDS-PAGE: Confirm that the correct reducing or non-reducing conditions were used; glycosylation or post-translational modifications introduced by the expression system may result in apparent molecular weights different from the theoretical value — consult the CoA for expected migration pattern.
• Loss of activity after storage: Ensure aliquots were stored at the correct temperature; avoid repeated freeze-thaw cycles; confirm that carrier protein (BSA) was added at reconstitution for long-term storage.

Frequently asked questions

What is the recommended reconstitution buffer for Recombinant Human GLP2R Protein?

It is generally recommended to reconstitute lyophilized recombinant proteins in sterile ultra-pure water or PBS (pH 7.4). For GLP2R specifically, consult the Certificate of Analysis (CoA) provided with lot rp370014 for the manufacturer-recommended reconstitution buffer and volume. Adding 0.1% BSA can help stabilize the reconstituted protein for storage.

How many freeze-thaw cycles can Recombinant Human GLP2R Protein tolerate?

To preserve protein integrity and biological activity, it is recommended to limit freeze-thaw cycles to a maximum of three. For best results, aliquot the reconstituted protein into single-use volumes immediately after reconstitution and store at -80°C to avoid repeated freeze-thaw.

What experimental applications is Recombinant Human GLP2R Protein validated for?

Recombinant Human GLP2R Protein (rp370014) is suitable for ligand binding assays, ELISA and immunoassay development, antibody screening and validation, drug discovery screening, and structural/biochemical studies including SPR and DSF. Please verify specific validated applications on the product CoA or contact Aladdin Scientific technical support for guidance.

Why does my GLP2R protein appear at an unexpected molecular weight on SDS-PAGE?

Apparent molecular weight on SDS-PAGE may differ from the theoretical value due to glycosylation, other post-translational modifications, or the presence of a purification tag introduced by the recombinant expression system. Consult the CoA supplied with rp370014 for the expected band size and any notes regarding protein modifications.

Is Recombinant Human GLP2R Protein suitable for in vivo or clinical use?

No. Recombinant Human GLP2R Protein (rp370014) is designated for Research Use Only (RUO). It is not intended, validated, or approved for therapeutic, diagnostic, or clinical applications in humans or animals.

References

1. Drucker DJ, Yusta B. Physiology and pharmacology of the enteroendocrine hormone glucagon-like peptide-2. Annu Rev Physiol. 2014;76:561–583. doi:10.1146/annurev-physiol-021113-170317
2. Munroe DG, Gupta A, Koller M, et al. Prototypic G-protein-coupled receptor for the intestinotrophic factor glucagon-like peptide 2. Proc Natl Acad Sci USA. 1999;96(4):1569–1573. PMID:9990065
3. Jeppesen PB, Gilroy R, Pertkiewicz M, et al. Randomised placebo-controlled trial of teduglutide in reducing parenteral nutrition and/or intravenous fluid requirements in patients with short bowel syndrome. Gut. 2011;60(7):902–914. doi:10.1136/gut.2010.218271

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