Recombinant Human ASXL3 Protein: Research Guide & Protocol

Recombinant Human ASXL3 Protein is a key epigenetic regulatory factor involved in chromatin remodeling and Polycomb-group complex function, offering researchers a validated tool for studying histone modification and gene silencing mechanisms.

Product Name

Recombinant Human ASXL3 Protein

SKU

rp376647

Protein Family

ASXL (Additional Sex Combs-Like) protein family

Biological Function

Chromatin remodeling and epigenetic regulation via Polycomb-group complexes

Species Origin

Human (Homo sapiens)

Primary Research Area

Epigenetics, chromatin biology, transcriptional regulation

Overview of ASXL3 and Its Biological Significance

ASXL3 (Additional Sex Combs-Like 3) is a member of the ASXL protein family, which plays essential roles in epigenetic regulation through its participation in Polycomb Repressive Complexes (PRCs). These complexes mediate histone H2A ubiquitination (H2AK119ub1) and contribute to stable gene silencing during development and differentiation.

In humans, the ASXL family comprises three members—ASXL1, ASXL2, and ASXL3—each exhibiting distinct tissue-specific expression patterns and associated phenotypes when mutated. ASXL3 mutations have been associated with Bainbridge–Ropers syndrome, a neurodevelopmental disorder characterized by intellectual disability and behavioral abnormalities, underscoring its importance in normal human development.

Recombinant Human ASXL3 Protein (rp376647) from Aladdin Scientific enables in vitro biochemical studies of Polycomb complex assembly, histone-binding assays, and protein–protein interaction analyses relevant to chromatin biology and disease research.

Reconstitution and Storage Guidelines

Proper reconstitution and storage of Recombinant Human ASXL3 Protein are critical for maintaining biological activity and reproducibility across experiments. Follow the guidelines below to ensure optimal performance:

• Reconstitution: Briefly centrifuge the vial before opening to bring lyophilized material to the bottom. Reconstitute in sterile deionized water or the buffer recommended in the specific product datasheet (typically PBS, pH 7.4) to a working concentration. Allow the solution to stand for 5–10 minutes at room temperature and mix gently; avoid vigorous vortexing.
• Working Aliquots: Prepare single-use aliquots immediately after reconstitution to minimize freeze–thaw cycles, which can reduce protein activity and integrity.
• Short-term Storage: Store reconstituted protein at 4 °C for up to 1 week if used frequently.
• Long-term Storage: For long-term storage, add a carrier protein (e.g., 0.1% BSA) if not already present, and store aliquots at −20 °C or −80 °C. Avoid repeated freeze–thaw cycles.
• Lyophilized Form: The lyophilized protein should be stored at −20 °C and is stable for up to 12 months from the date of manufacture under proper conditions.

Note: Always consult the product-specific datasheet (absea#PP-10190) for exact buffer composition, concentration, and storage conditions, as formulations may vary by lot.

Experimental Applications and Protocols

Recombinant Human ASXL3 Protein is suitable for a range of in vitro biochemical and cell-free assay formats. The following applications represent common uses supported by the recombinant protein format:

1. Protein–Protein Interaction Studies: Use in pull-down assays or co-immunoprecipitation (co-IP) setups to characterize ASXL3 interactions with BAP1, BRCA1-Associated Protein 1, and other Polycomb complex components. Typical input concentrations range from 0.5–2 µg per assay; optimize empirically.
2. Histone-Binding Assays: Employ in histone peptide pulldown assays to investigate ASXL3 PHD finger domain recognition of histone H3 modifications. Use biotinylated histone peptides on streptavidin beads with 1–5 µg recombinant protein per reaction.
3. Enzyme Activity Assays: As a component of the PR-DUB (Polycomb Repressive DeUBiquitinase) complex, ASXL3 can be used to reconstitute deubiquitinase activity together with BAP1 in vitro. Validate activity using ubiquitinated H2A substrate and western blotting or fluorescence-based readouts.
4. Western Blot Positive Control: Use as a positive control to validate anti-ASXL3 antibody performance. Load 50–100 ng per lane on SDS-PAGE followed by standard immunoblotting protocols.
5. ELISA Standard Curve: Prepare serial dilutions of the recombinant protein for use as a standard in sandwich ELISA for quantification of endogenous ASXL3 levels in biological samples.

Optimize all conditions empirically. Consult the product datasheet for guaranteed tested applications specific to lot rp376647.

Quality, Purity, and Safety Considerations

When working with Recombinant Human ASXL3 Protein, researchers should be aware of the following quality and safety considerations to ensure valid experimental results and laboratory safety:

• Purity: Recombinant proteins intended for research use are typically assessed by SDS-PAGE and/or HPLC. Verify lot-specific purity data on the Certificate of Analysis (CoA) supplied with each shipment. Flag any purity below the stated specification before use.
• Endotoxin Levels: For cell-based assays, endotoxin contamination can confound results. Check the CoA for LAL (Limulus Amebocyte Lysate) endotoxin test values. If endotoxin levels are a concern for your assay, contact Aladdin Scientific technical support for low-endotoxin options.
• Biological Safety: Recombinant Human ASXL3 Protein is a research-use-only (RUO) reagent and is not intended for diagnostic or therapeutic use in humans or animals. Handle according to your institution's biosafety guidelines for recombinant proteins. Use appropriate personal protective equipment (PPE) including gloves and lab coat.
• Carrier Proteins and Additives: The formulation may contain BSA or glycerol as stabilizers. Ensure these components are compatible with your downstream assay before use.
• Lot-to-Lot Consistency: For longitudinal studies, order sufficient stock from a single lot where possible to minimize variability. Contact Aladdin Scientific procurement for lot reservation options.

Frequently asked questions

What is the recommended reconstitution buffer for Recombinant Human ASXL3 Protein?

The protein should be reconstituted in sterile deionized water or PBS (pH 7.4) as generally recommended for recombinant proteins of this class. Always refer to the lot-specific product datasheet (absea#PP-10190) for the exact recommended buffer, concentration, and any stabilizing additives, as formulations may vary between lots.

How many freeze–thaw cycles can Recombinant Human ASXL3 Protein withstand without significant loss of activity?

As a general best practice for recombinant proteins, repeated freeze–thaw cycles should be minimized—ideally limited to three or fewer—to preserve protein integrity and activity. We recommend preparing single-use aliquots immediately after reconstitution. Please confirm lot-specific stability data with Aladdin Scientific technical support or refer to the product CoA.

Is Recombinant Human ASXL3 Protein suitable for use as a Western blot positive control?

Yes, Recombinant Human ASXL3 Protein (rp376647) can be used as a positive control to validate the performance of anti-ASXL3 antibodies in Western blotting. A loading amount of 50–100 ng per lane on SDS-PAGE is typically sufficient, though optimal amounts should be determined empirically for your specific antibody and detection system.

What are the primary biological functions of ASXL3 relevant to my research?

ASXL3 functions as an epigenetic regulator within Polycomb Repressive Complexes, contributing to histone H2A deubiquitination as part of the PR-DUB complex with BAP1, and to gene silencing through chromatin remodeling. It has been implicated in neurodevelopmental disorders (Bainbridge–Ropers syndrome) and is a subject of active research in developmental biology and cancer epigenetics.

Can I use this recombinant protein for diagnostic or therapeutic purposes?

No. Recombinant Human ASXL3 Protein (rp376647) is a research-use-only (RUO) reagent and is strictly not intended for diagnostic, therapeutic, or clinical use in humans or animals. It is designed exclusively for in vitro laboratory research applications.

References

1. Bainbridge MN et al. De novo truncating mutations in ASXL3 are associated with a novel clinical phenotype with similarities to Bohring-Opitz syndrome. Genome Medicine. 2013.
2. Abdel-Wahab O, Dey A. The ASXL-BAP1 axis: new factors in myelopoiesis, cancer and epigenetics. Leukemia. 2013.
3. Campagne A et al. BAP1 complex promotes transcription by opposing PRC1-mediated H2A ubiquitylation. Nature Communications. 2019.

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