Recombinant Human L3MBTL1 Protein: Research Guidelines

Recombinant Human L3MBTL1 Protein is a chromatin-associated epigenetic reader protein produced recombinantly for use in histone methylation binding studies, chromatin regulation research, and related biochemical assays.

Product Name

Recombinant Human L3MBTL1 Protein

SKU

rp376604

Protein Family

MBT (malignant brain tumor) domain-containing protein family

Biological Function

Histone methyl-lysine reader; chromatin compaction and transcriptional repression

Primary Research Area

Epigenetics, chromatin biology, cancer biology

Recommended Storage

-80°C for long-term storage; avoid repeated freeze-thaw cycles

Product Overview and Biological Background

L3MBTL1 (Lethal(3) Malignant Brain Tumor-Like Protein 1) is a member of the MBT domain-containing protein family, which plays a critical role in epigenetic regulation. The protein contains three MBT repeat domains that function as readers of mono- and di-methylated histone lysine residues, particularly histone H4 lysine 20 (H4K20me1/me2) and histone H1b lysine 26. Through these interactions, L3MBTL1 contributes to chromatin compaction and transcriptional repression, acting as a component of diverse chromatin-modifying complexes.

Recombinant Human L3MBTL1 Protein (SKU: rp376604) is produced to support in vitro biochemical studies of histone-reader interactions, chromatin compaction mechanisms, and the broader regulatory networks involving Polycomb-group proteins. L3MBTL1 has also been implicated in tumor suppression, DNA damage response, and cell-cycle regulation, making it a relevant target in cancer biology research.

Reconstitution and Storage Protocol

Reconstitution: Upon receipt, briefly centrifuge the vial before opening to consolidate the lyophilized material (if supplied lyophilized). Reconstitute in an appropriate sterile buffer such as phosphate-buffered saline (PBS, pH 7.4) containing a carrier protein (e.g., 0.1% BSA) to prevent adsorption to tube surfaces. Gently mix by pipetting; do not vortex vigorously, as this may denature the protein. Refer to the specific product datasheet for the recommended reconstitution volume and concentration.

Storage conditions: Aliquot reconstituted protein into single-use volumes to avoid repeated freeze-thaw cycles. Store aliquots at -80°C for long-term storage or at -20°C for short-term use (up to 1 month). Once thawed, keep on ice during experiments and do not refreeze. Stability may vary depending on buffer composition and downstream application; always consult the product datasheet for lot-specific recommendations.

Shipping: The product is typically shipped on dry ice. Upon receipt, immediately transfer to the appropriate storage temperature and record the receipt date on the vial for traceability.

Experimental Applications and Usage Guidelines

Recombinant Human L3MBTL1 Protein is suitable for a range of biochemical and cell-free assays, including but not limited to:

• Histone peptide pull-down assays: L3MBTL1 can be used to investigate binding affinity and specificity toward mono- and di-methylated histone lysine peptides (e.g., H4K20me1, H4K20me2).
• Protein–protein interaction studies: Co-immunoprecipitation (Co-IP) or GST pull-down experiments can be performed to map interaction partners within chromatin-modifying complexes.
• Electrophoretic mobility shift assays (EMSA): To assess nucleosome or DNA binding properties of L3MBTL1 under defined conditions.
• Enzyme-linked immunosorbent assays (ELISA) and surface plasmon resonance (SPR): For quantitative binding kinetics between L3MBTL1 and histone substrates or small-molecule inhibitors.
• Structural biology: As a component in crystallography or cryo-EM sample preparation to elucidate MBT domain–histone interactions at atomic resolution.

When designing experiments, include appropriate positive and negative controls, and optimize protein concentration empirically for each assay format. Always keep the protein on ice when in use, and minimize handling time at room temperature.

Quality, Purity, and Safety Considerations

Purity and quality: Recombinant Human L3MBTL1 Protein (rp376604) is manufactured under controlled conditions to ensure consistency between lots. Purity is typically assessed by SDS-PAGE and/or HPLC analysis. Endotoxin levels are measured to meet research-grade standards. Specific purity values, endotoxin thresholds, and activity data are provided on the lot-specific Certificate of Analysis (CoA), which is available upon request from Aladdin Scientific.

Biological safety: Although this recombinant protein is not classified as a biohazard under standard laboratory regulations, researchers should follow institutional biosafety guidelines and standard laboratory best practices when handling any recombinant protein. Use appropriate personal protective equipment (PPE) including gloves and lab coat. Avoid skin and eye contact.

Disposal: Dispose of protein solutions and contaminated materials according to your institution's chemical and biological waste disposal regulations.

Frequently asked questions

What is the recommended buffer for reconstituting Recombinant Human L3MBTL1 Protein?

A sterile phosphate-buffered saline (PBS, pH 7.4) supplemented with 0.1% BSA is generally recommended as a reconstitution buffer to maintain protein stability and prevent adsorption to tube walls. Always consult the lot-specific datasheet or Certificate of Analysis for the exact recommended conditions.

How should I store Recombinant Human L3MBTL1 Protein to maximize stability?

After reconstitution, aliquot the protein into single-use volumes and store at -80°C for long-term stability. Avoid repeated freeze-thaw cycles, as these can lead to protein aggregation and loss of activity. For experiments, thaw only the required aliquot and keep on ice throughout use.

What experimental applications is Recombinant Human L3MBTL1 Protein validated for?

Recombinant Human L3MBTL1 Protein is suitable for histone peptide pull-down assays, protein–protein interaction studies (Co-IP, GST pull-down), EMSA, ELISA, surface plasmon resonance (SPR), and structural biology applications. Specific validated applications for lot rp376604 are detailed in the product datasheet available from Aladdin Scientific.

What is the biological significance of L3MBTL1 in epigenetic regulation?

L3MBTL1 functions as an epigenetic reader that recognizes mono- and di-methylated histone lysine marks, particularly H4K20me1 and H4K20me2, via its MBT repeat domains. This recognition promotes chromatin compaction and transcriptional repression. L3MBTL1 has also been linked to tumor suppression and DNA damage response pathways, making it relevant to both basic epigenetics research and cancer biology.

References

1. Trojer P, et al. L3MBTL1, a histone-methylation-dependent chromatin lock. Cell. 2007;129(5):915-928. doi:10.1016/j.cell.2007.03.048
2. Min J, et al. Structural basis for specific binding of Polycomb chromodomain to histone H3 methylated at Lys 27. Genes Dev. 2003;17(15):1823-1828. doi:10.1101/gad.269603
3. Kalakonda N, et al. Histone H4 lysine 20 monomethylation promotes transcriptional repression by L3MBTL1. Oncogene. 2008;27(31):4293-4304. doi:10.1038/onc.2008.67

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