Recombinant Human LCORL Protein: Protocol & Usage Guide

Recombinant Human LCORL Protein is a research-grade recombinant protein designed to support studies of transcriptional regulation, chromatin binding, and related molecular biology applications.

Product Name

Recombinant Human LCORL Protein

SKU

rp366951

Protein Origin

Human (Homo sapiens)

Protein Type

Recombinant protein, transcriptional regulator

Primary Application

Transcriptional regulation and chromatin biology research

Storage Recommendation

Avoid repeated freeze-thaw cycles; store according to product datasheet

Overview of LCORL and Its Biological Significance

LCORL (Ligand-Dependent Nuclear Receptor Corepressor-Like) is a transcriptional repressor protein encoded by the LCORL gene in humans. It belongs to a family of proteins characterized by a BTB/POZ domain and plays important roles in regulating gene expression, chromatin organization, and developmental processes.

Research has implicated LCORL in growth regulation, with genome-wide association studies (GWAS) identifying variants near the LCORL locus associated with human stature and body size traits. Understanding LCORL protein function at the molecular level requires high-quality recombinant protein reagents suitable for biochemical and cell-based assays.

Reconstitution and Preparation Protocol

Follow these general steps when reconstituting Recombinant Human LCORL Protein (rp366951). Always consult the product-specific datasheet for exact buffer compositions, recommended concentrations, and any product-specific instructions before proceeding.

1. Equilibrate to room temperature: Allow the vial to equilibrate to room temperature for 15–30 minutes before opening to minimize condensation and protein aggregation.
2. Prepare reconstitution buffer: Use the buffer specified in the product datasheet (typically sterile PBS, pH 7.4, or a buffer containing a carrier protein such as 0.1% BSA) to maximize solubility and stability.
3. Add reconstitution buffer gently: Add the recommended volume of buffer slowly along the side of the vial. Do not pipette directly onto the lyophilized cake to avoid foaming and denaturation.
4. Mix gently: Swirl or gently rotate the vial to dissolve the protein. Do not vortex, as this may cause denaturation.
5. Centrifuge briefly: Spin down at low speed (e.g., 2,000–5,000 × g for 1 minute) to collect any residual material from the vial walls.
6. Aliquot immediately: Divide the reconstituted protein into single-use aliquots appropriate to your experimental needs to avoid repeated freeze-thaw cycles.

Storage, Stability, and Handling Guidelines

Proper storage and handling of Recombinant Human LCORL Protein are critical to maintaining protein integrity and experimental reproducibility. Adhere to the following best practices:

• Long-term storage: Store lyophilized protein at –20°C or –80°C as specified on the product datasheet. Reconstituted protein should typically be stored at –80°C for long-term use.
• Avoid freeze-thaw cycles: Repeated freeze-thaw cycles significantly reduce protein activity. Single-use aliquots are strongly recommended.
• Working aliquots: For short-term use (≤1 week), reconstituted protein may be stored at 4°C; verify acceptable duration in the product datasheet.
• Carrier protein: Adding a carrier protein (e.g., 0.1% BSA) to the reconstitution buffer may improve stability and reduce adsorption to tube surfaces.
• Avoid detergents and extreme pH: Unless required by your assay, avoid exposing the protein to strong detergents, extreme pH, or high temperatures that may cause denaturation.
• Expiry and lot-specific information: Check lot-specific Certificate of Analysis (CoA) for purity, concentration, and expiry date before use.

Recommended Applications and Experimental Tips

Recombinant Human LCORL Protein (rp366951) is suitable for a range of research applications. Optimal conditions may vary by assay; pilot experiments are recommended to determine best working concentrations and conditions.

• Protein–protein interaction studies: LCORL may be used in pull-down assays or co-immunoprecipitation (co-IP) experiments to identify interacting partners involved in transcriptional repression complexes.
• Biochemical assays: The recombinant protein can be employed in ELISA, Western blot validation of antibodies, or as a standard for quantification purposes.
• Structural and biophysical studies: Suitable for use in techniques such as surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), or circular dichroism (CD) to characterize binding kinetics and protein folding.
• Cell-free transcription assays: May be used in in vitro transcription systems to study the repressor activity of LCORL on target gene promoters.

Tip: Always include a negative control (e.g., buffer alone or an irrelevant protein of similar molecular weight) to validate specificity in functional assays. Confirm protein activity using a validated functional assay before critical experiments.

Quality Control and Troubleshooting

To ensure reliable experimental outcomes, implement the following quality control (QC) measures and refer to this troubleshooting guide if problems arise.

Quality Control Checks

• Verify protein identity and purity by SDS-PAGE prior to use; expected purity should meet the specification stated on the CoA.
• Confirm protein concentration using a validated colorimetric assay (e.g., BCA or Bradford assay) against a BSA standard curve.
• Check for aggregation by visual inspection or dynamic light scattering (DLS) if critical for your application.

Common Issues and Solutions

• Protein precipitation or aggregation: Ensure reconstitution buffer is appropriate and at the correct pH. Work at 4°C when possible and avoid vortexing. Consider adding a mild detergent or increasing salt concentration if recommended by the datasheet.
• Low protein activity: Confirm the protein has not been subjected to excessive freeze-thaw cycles. Verify storage conditions and check expiry date. Ensure assay conditions (pH, temperature, cofactors) are appropriate for LCORL function.
• Unexpected band pattern on SDS-PAGE: A minor band in addition to the main band may represent post-translational modifications, proteolytic fragments, or protein dimers. Refer to the CoA for expected molecular weight and purity profile.

Frequently asked questions

What is LCORL and why is it important for research?

LCORL (Ligand-Dependent Nuclear Receptor Corepressor-Like) is a transcriptional repressor protein involved in regulating gene expression and chromatin organization. It has been associated with growth regulation and body size traits in genome-wide association studies, making it of interest in developmental biology, genetics, and disease research.

How should I reconstitute Recombinant Human LCORL Protein?

Allow the vial to equilibrate to room temperature before opening. Add the recommended reconstitution buffer (see product datasheet) gently along the vial wall, swirl to mix without vortexing, and centrifuge briefly. Aliquot immediately into single-use portions and store as directed to avoid freeze-thaw degradation.

What applications is Recombinant Human LCORL Protein (rp366951) validated for?

Recombinant Human LCORL Protein is suitable for use in protein–protein interaction studies, ELISA, Western blot antibody validation, structural and biophysical characterization, and in vitro transcription assays. Please consult the product datasheet for the specific validated applications and recommended conditions for this lot.

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

As with most recombinant proteins, repeated freeze-thaw cycles are not recommended, as they can degrade protein integrity and reduce activity. It is best practice to aliquot the protein into single-use volumes upon first reconstitution to minimize freeze-thaw exposure. Refer to the product datasheet for lot-specific stability information.

Where can I find the Certificate of Analysis (CoA) for rp366951?

The Certificate of Analysis for Recombinant Human LCORL Protein (rp366951) can be obtained from Aladdin Scientific's website by entering the product SKU or lot number, or by contacting Aladdin Scientific customer service directly. The CoA contains lot-specific purity, concentration, and quality data.

References

1. Weedon MN, et al. Genome-wide association analysis identifies 20 loci that influence adult height. Nature Genetics. 2008.
2. Gudbjartsson DF, et al. Many sequence variants affecting diversity of adult human height. Nature Genetics. 2008.
3. Aladdin Scientific. Recombinant Human LCORL Protein (rp366951) Product Datasheet. Aladdin Scientific.

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