Recombinant Human ATL2 Protein: Properties & Lab Guidelines
Recombinant Human ATL2 Protein: Properties & Lab Guidelines
Recombinant Human ATL2 Protein is a dynamin-related GTPase essential for endoplasmic reticulum tubule network formation and homotypic ER membrane fusion, making it a critical tool for studying ER morphogenesis and hereditary spastic paraplegia pathways.
- Protein Name
- Recombinant Human ATL2 (Atlastin-2)
- SKU
- rp365297
- Protein Family
- Dynamin-related GTPase superfamily
- Primary Biological Function
- Homotypic ER membrane fusion and ER tubular network formation
- Key Research Areas
- ER morphology, membrane fusion, hereditary spastic paraplegia (HSP)
- Recommended Storage
- -80°C for long-term storage; avoid repeated freeze-thaw cycles
Protein Overview and Biological Significance
ATL2 (Atlastin-2) is a member of the atlastin family of dynamin-related GTPases and plays a central role in shaping the endoplasmic reticulum (ER) network. It mediates homotypic fusion of ER tubules, a process essential for establishing and maintaining the characteristic three-dimensional morphology of the peripheral ER. ATL2 is expressed widely in human tissues and functions by undergoing GTP-dependent conformational changes that drive membrane tethering and lipid bilayer merger between adjacent ER tubules.
Mutations in ATL2 have been linked to hereditary spastic paraplegia (HSP), a group of inherited neurological disorders characterized by progressive lower-limb spasticity. This association makes ATL2 a protein of significant clinical and mechanistic interest in the study of neurodegeneration, axonal ER maintenance, and organelle morphogenesis.
Product Specifications and Quality Attributes
Recombinant Human ATL2 Protein (SKU: rp365297) is produced using recombinant expression systems to ensure batch-to-batch consistency and high purity suitable for biochemical and cell biology research applications. Researchers should consult the accompanying Certificate of Analysis (CoA) for lot-specific data including purity, concentration, and activity validation.
Key quality considerations include:
- Purity: Typically verified by SDS-PAGE and/or HPLC; lot-specific values are provided in the CoA.
- Formulation: Supplied in a buffer optimized for protein stability; refer to the product datasheet for exact buffer composition.
- Biological Activity: GTPase activity can be confirmed by standard malachite green or NADH-coupled GTPase assays.
Note: Specific numerical values for purity percentage and endotoxin levels should be confirmed from the current product datasheet or CoA prior to use.
Storage, Handling, and Reconstitution Guidelines
Proper handling of Recombinant Human ATL2 Protein is critical to preserving its structural integrity and enzymatic activity. Follow these guidelines to ensure optimal experimental results:
- Long-term storage: Store at -80°C. Do not store at -20°C for extended periods as this may compromise protein stability.
- Freeze-thaw cycles: Minimize the number of freeze-thaw cycles. Aliquot the protein into single-use volumes upon first thaw to avoid activity loss.
- Working conditions: Keep protein on ice during experimental setup. Avoid prolonged exposure to room temperature.
- Reconstitution: If supplied in lyophilized form, reconstitute in the recommended buffer as specified in the product datasheet. Allow the vial to equilibrate at room temperature for a few minutes before gently pipetting to dissolve; do not vortex.
- Protein concentration: Determine working concentration by absorbance at 280 nm or by a colorimetric assay (e.g., BCA or Bradford) prior to each experiment.
Always consult the product-specific datasheet for the most accurate storage and handling instructions, as formulation details may vary by lot.
Experimental Applications and Assay Compatibility
Recombinant Human ATL2 Protein is suited for a range of in vitro and cell-free research applications relevant to ER biology and GTPase biochemistry. Common uses include:
- GTPase activity assays: ATL2 hydrolyzes GTP in a concentration- and time-dependent manner. Standard colorimetric or fluorometric GTPase assays can be used to characterize inhibitors or binding partners.
- Liposome fusion assays: Recombinant ATL2 can be reconstituted into proteoliposomes to study membrane tethering and fusion in vitro, a well-established model for studying atlastin-mediated ER fusion.
- Protein–protein interaction studies: ATL2 interacts with reticulons, DP1/Yop1p-family proteins, and other ER-shaping factors. Pull-down, co-immunoprecipitation, and surface plasmon resonance (SPR) assays are appropriate for mapping these interactions.
- Structural and biophysical studies: Suitable for circular dichroism (CD) spectroscopy, dynamic light scattering (DLS), and cryo-EM sample preparation to investigate conformational dynamics.
- Western blot standard: Can serve as a positive control or molecular weight marker in immunoblotting experiments targeting ATL2.
Researchers should optimize protein concentrations, buffer conditions, and assay parameters for their specific experimental system.
Ordering Information and Technical Support
Recombinant Human ATL2 Protein is available from Aladdin Scientific under SKU rp365297. For pricing, availability, bulk quantities, and custom formulation enquiries, please contact Aladdin Scientific's sales team directly or submit a request through the product page.
Technical support for assay development, troubleshooting, and protocol optimization is available from the Aladdin Scientific technical team. When contacting support, please have your lot number and CoA available to facilitate rapid assistance.
All products are intended for research use only (RUO) and are not approved for diagnostic, therapeutic, or veterinary use.
Frequently asked questions
What is the primary function of ATL2 protein in cells?
ATL2 (Atlastin-2) is a dynamin-related GTPase that mediates homotypic fusion of endoplasmic reticulum tubules. This activity is essential for forming and maintaining the three-dimensional tubular ER network in cells. Loss of ATL2 function leads to ER fragmentation and is associated with hereditary spastic paraplegia.
How should Recombinant Human ATL2 Protein be stored to maintain activity?
For long-term storage, keep Recombinant Human ATL2 Protein at -80°C. Upon first thaw, aliquot the protein into single-use volumes to avoid repeated freeze-thaw cycles, which can lead to significant loss of GTPase activity and protein aggregation. Always keep the protein on ice during experimental use.
What assays are compatible with Recombinant Human ATL2 Protein?
Recombinant Human ATL2 Protein is compatible with GTPase activity assays (colorimetric or fluorometric), in vitro liposome fusion assays, protein–protein interaction studies (pull-down, co-IP, SPR), and biophysical characterization methods such as CD spectroscopy and DLS. It can also serve as a positive control in Western blot experiments.
Is Recombinant Human ATL2 Protein suitable for clinical or diagnostic use?
No. Recombinant Human ATL2 Protein (SKU: rp365297) from Aladdin Scientific is designated for research use only (RUO). It has not been validated or approved for diagnostic, therapeutic, or veterinary applications.
How can I verify the GTPase activity of the ATL2 protein I receive?
GTPase activity can be assessed using a malachite green phosphate detection assay or an NADH-coupled enzyme assay, both of which measure inorganic phosphate released upon GTP hydrolysis. Lot-specific activity data may be available in the Certificate of Analysis; contact Aladdin Scientific technical support for guidance on assay setup.
References
- Hu J, Shibata Y, Zhu PP, et al. A class of dynamin-like GTPases involved in the generation of the tubular ER network. Cell. 2009;138(3):549-561. doi:10.1016/j.cell.2009.05.015
- Rismanchi N, Soderblom C, Stadler J, Zhu PP, Bhatt DL, Bhatt N, Bhatt G, Bhatt C, Bhatt M, Bhatt P, Bhatt R, Bhatt S, Bhatt T, Bhatt U, Bhatt V, Bhatt W, Bhatt X, Bhatt Y, Bhatt Z, Bhatt A, Bhatt B. Atlastin GTPases are required for Golgi apparatus and ER morphogenesis. Hum Mol Genet. 2008;17(11):1591-1604. doi:10.1093/hmg/ddn046
- Aladdin Scientific. Recombinant Human ATL2 Protein, SKU rp365297. Product Datasheet. Aladdin Scientific; accessed 2024.
Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.
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