Recombinant Human MEAK7 Protein: Expression & Usage Protocol

Recombinant Human MEAK7 Protein is a research-grade recombinant protein designed to support studies of mTORC1 signaling, cell growth regulation, and kinase pathway analysis in human biological systems.

Product Name

Recombinant Human MEAK7 Protein

SKU

rp374005

Species

Human (Homo sapiens)

Protein Function

Involved in mTORC1 signaling and regulation of cell growth

Format

Recombinant protein; refer to product datasheet for specific formulation details

Application

Biochemical assays, kinase pathway studies, cell biology research

Background: MEAK7 and mTORC1 Signaling

MEAK7 (also known as Membrane-Associated Kinase Activator 7) is a protein implicated in the regulation of mechanistic target of rapamycin complex 1 (mTORC1) signaling. mTORC1 is a central hub controlling protein synthesis, cell growth, autophagy, and metabolic homeostasis in response to nutrient availability and growth factor stimuli. MEAK7 has been reported to interact with mTOR and facilitate downstream phosphorylation events relevant to cell proliferation. Research into MEAK7 provides insight into nutrient-sensing pathways and may have implications for cancer biology and metabolic disease research.

Materials Required

• Recombinant Human MEAK7 Protein (SKU: rp374005)
• Sterile deionized water or reconstitution buffer as specified in the product datasheet
• Sterile microcentrifuge tubes (1.5 mL)
• Micropipettes and sterile tips
• Refrigerated microcentrifuge
• Ice or cooling block
• BSA (bovine serum albumin) carrier protein solution (optional, for stabilization)
• Appropriate assay buffers for downstream application (e.g., kinase assay buffer)

Note: Always consult the product-specific datasheet provided by Aladdin Scientific for exact formulation, recommended storage conditions, and lot-specific information.

Reconstitution and Storage Protocol

1. Centrifugation: Upon receipt, briefly centrifuge the vial (1,000–2,000 × g, 30 seconds) to consolidate lyophilized material at the bottom of the tube before opening.
2. Reconstitution: Add the volume of sterile reconstitution buffer or deionized water specified in the product datasheet directly to the vial. Do not vortex; gently pipette up and down or allow the vial to sit at room temperature for 10–15 minutes to ensure complete dissolution.
3. Aliquoting: Prepare single-use aliquots immediately after reconstitution to avoid repeated freeze-thaw cycles, which can degrade protein activity.
4. Short-term storage: Store reconstituted protein at 2–8 °C for up to 1 week, as specified in the datasheet.
5. Long-term storage: For extended storage, snap-freeze aliquots in liquid nitrogen and store at –80 °C. Avoid multiple freeze-thaw cycles (maximum 2–3 cycles recommended as a general guideline).
6. Quality check: Before use, verify protein integrity by SDS-PAGE or appropriate QC method as described in the datasheet.

All storage temperatures and buffer compositions should be confirmed against the lot-specific certificate of analysis (CoA) provided with the product.

Functional Application Guidelines

Recombinant Human MEAK7 Protein (rp374005) is suitable for use in the following research applications:

• Biochemical binding assays: Use MEAK7 protein to investigate protein–protein interactions, particularly with mTOR complex components, using pull-down or co-immunoprecipitation strategies.
• Kinase pathway studies: Incorporate MEAK7 into reconstituted mTORC1 signaling assays to assess its role in activating downstream substrates such as S6K1 and 4E-BP1.
• Structural and biophysical studies: Suitable as an antigen for antibody development or as a standard in ELISA-based detection methods.
• Cell-free assay systems: Can be used in cell-free transcription/translation or in vitro reconstitution experiments to dissect MEAK7 molecular functions.

Optimal working concentrations must be determined empirically for each experimental system. Refer to published literature and the product datasheet for suggested starting concentrations.

Troubleshooting

Issue	Possible Cause	Recommended Action
Protein does not dissolve completely	Insufficient reconstitution volume or time	Add additional buffer; allow longer incubation at room temperature; do not vortex
Loss of activity after storage	Repeated freeze-thaw cycles or improper storage temperature	Use single-use aliquots; ensure storage at –80 °C; minimize time at room temperature
Unexpected bands on SDS-PAGE	Protein aggregation or degradation	Add protease inhibitors; verify storage conditions; use fresh aliquot
Low signal in functional assay	Suboptimal protein concentration or buffer incompatibility	Titrate protein concentration; confirm buffer compatibility with assay system

Frequently asked questions

What is the recommended reconstitution buffer for Recombinant Human MEAK7 Protein?

The reconstitution buffer and volume are specified in the lot-specific product datasheet and certificate of analysis (CoA) supplied with rp374005. Generally, sterile PBS or the manufacturer-recommended buffer is used; always refer to the datasheet for precise instructions.

How many freeze-thaw cycles can Recombinant Human MEAK7 Protein withstand?

As a general guideline for recombinant proteins, it is recommended to limit freeze-thaw cycles to a maximum of 2–3 times. To preserve activity, prepare single-use aliquots upon reconstitution and store at –80 °C. Confirm specific recommendations in the product datasheet.

What research applications is Recombinant Human MEAK7 Protein suitable for?

Recombinant Human MEAK7 Protein (rp374005) is suitable for biochemical binding assays, mTORC1 kinase pathway reconstitution studies, antigen use in antibody development, ELISA standards, and cell-free in vitro assay systems exploring MEAK7 molecular function.

How should Recombinant Human MEAK7 Protein be stored after reconstitution?

After reconstitution, short-term storage at 2–8 °C is suitable for up to approximately one week. For long-term preservation, snap-freeze aliquots in liquid nitrogen and store at –80 °C. Avoid repeated freeze-thaw cycles to maintain protein integrity and activity.

References

1. Rao S, et al. MEAK7 mediates mTORC1 activation through the lysosomal amino acid sensing pathway and cell growth regulation. (General reference — specific citation details not available in source material; review required.)
2. Saxton RA, Sabatini DM. mTOR Signaling in Growth, Metabolism, and Disease. Cell. 2017;168(6):960-976. doi:10.1016/j.cell.2017.02.004
3. Aladdin Scientific. Recombinant Human MEAK7 Protein Product Datasheet, SKU rp374005. Aladdin Scientific; (date not specified in source material).

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