Recombinant Human GORAB Protein: Production & Usage Protocol

Recombinant Human GORAB Protein is a research-grade recombinant protein targeting the Golgi RAB GTPase-binding protein GORAB, supplied by Aladdin Scientific (SKU: rp373957) to support studies in Golgi apparatus function, vesicular trafficking, and the molecular basis of craniofacial and skeletal dysplasia disorders.

Product Name

Recombinant Human GORAB Protein

SKU

rp373957

Source Organism

Human (Homo sapiens)

Protein Target

GORAB (Golgi RAB GTPase-binding protein; also known as Scyl1BP1)

Primary Research Applications

Protein interaction studies, functional assays, Western blot, ELISA, cell biology research

Relevant Disease Area

Geroderma osteodysplastica, craniofacial and skeletal dysplasia

Background: GORAB Protein Biology

GORAB (Golgi RAB GTPase-binding protein), also referred to as Scyl1BP1, is a protein encoded by the GORAB gene in humans. It localizes predominantly to the trans-Golgi network and plays an essential role in Golgi apparatus homeostasis and vesicular trafficking. GORAB interacts with RAB6 and ARF4 GTPases, positioning it as a critical scaffold protein in the coordination of Golgi-associated membrane transport.

Loss-of-function mutations in GORAB are causatively linked to Geroderma osteodysplastica (GO), a rare autosomal recessive connective tissue disorder characterized by wrinkled skin, osteoporosis, and craniofacial abnormalities. Research into GORAB function therefore has direct relevance to understanding Golgi biology, extracellular matrix secretion, and the pathogenesis of skeletal dysplasia syndromes.

Reconstitution Protocol

Follow these steps to reconstitute Recombinant Human GORAB Protein (rp373957) for experimental use:

1. Equilibration: Allow the vial to equilibrate to room temperature for 10–15 minutes before opening. Do not expose the lyophilized protein to direct heat.
2. Reconstitution buffer: Reconstitute the lyophilized protein in sterile, deionized water or an appropriate buffer (e.g., PBS, pH 7.4) to obtain the desired working concentration. Gently swirl the vial; do not vortex vigorously.
3. Centrifugation: Briefly centrifuge the vial (e.g., 5–10 seconds at low speed) prior to opening to collect any lyophilized material from the cap and walls.
4. Complete dissolution: Allow the solution to stand for 5 minutes at room temperature after addition of buffer to ensure complete dissolution before use.

Note: Refer to the product datasheet for the specific recommended reconstitution volume and concentration, as these values are lot-specific.

Storage and Stability Guidelines

Proper storage is essential to maintain the activity and integrity of Recombinant Human GORAB Protein:

• Lyophilized form: Store at –20°C or –80°C upon receipt. Avoid repeated freeze–thaw cycles, as these can compromise protein integrity and activity.
• Reconstituted protein: For short-term use (up to 1 week), store aliquots at 4°C. For longer-term storage, prepare single-use aliquots and store at –80°C.
• Carrier protein: Addition of a carrier protein such as 0.1% BSA (bovine serum albumin) to the reconstitution buffer may enhance stability, particularly at low protein concentrations. Verify compatibility with your downstream assay before use.
• Avoid: Repeated freeze–thaw cycles, prolonged exposure to room temperature, and storage in frost-free freezers that undergo temperature cycling.

Refer to the product-specific datasheet for confirmed stability data and recommended storage conditions for lot rp373957.

Experimental Applications

Recombinant Human GORAB Protein (rp373957) is suitable for a range of in vitro research applications, including but not limited to:

• Protein–protein interaction studies: Use in pull-down assays, co-immunoprecipitation validation, or surface plasmon resonance (SPR) to characterize GORAB interactions with RAB GTPases or other binding partners.
• Western blot (WB) standard: Use as a positive control or molecular weight standard to validate anti-GORAB antibodies in immunoblotting experiments.
• ELISA development: Serve as a standard or capture antigen in sandwich or competitive ELISA formats for quantification of GORAB in biological samples.
• Functional/enzymatic assays: Investigate GORAB's role in Golgi trafficking reconstitution systems or cell-free assays where purified components are required.
• Antibody production and validation: Use as an immunogen or validation reagent for generating and testing anti-GORAB antibodies.

Optimization of protein concentration, buffer conditions, and assay format may be required depending on the specific experimental system employed.

Quality, Safety, and Handling Considerations

Quality: Recombinant Human GORAB Protein from Aladdin Scientific is produced under controlled conditions with quality-control testing. Purity, concentration, and activity data are provided on the lot-specific Certificate of Analysis (CoA). Researchers should consult the CoA for confirmed specifications before use.

Safety: This product is intended for research use only (RUO) and is not approved for diagnostic or therapeutic use in humans or animals. Handle using standard laboratory safety practices. Wear appropriate personal protective equipment (PPE) including gloves and a laboratory coat. Although this recombinant protein is not classified as hazardous under standard laboratory conditions, follow your institution's biosafety guidelines.

Handling: Minimize exposure to room temperature during handling. Use sterile techniques when preparing aliquots to prevent microbial contamination, which can degrade protein quality. Dispose of any unused material in accordance with your institution's waste disposal regulations.

Frequently asked questions

What is GORAB protein and why is it important in research?

GORAB (Golgi RAB GTPase-binding protein) is a scaffold protein localized to the trans-Golgi network, where it coordinates vesicular trafficking by interacting with RAB6 and ARF4 GTPases. It is important because loss-of-function mutations in GORAB cause Geroderma osteodysplastica, a rare skeletal and connective tissue disorder, making it a key target for studies in Golgi biology and rare disease mechanisms.

How should I reconstitute Recombinant Human GORAB Protein (rp373957)?

Allow the vial to equilibrate to room temperature before opening, then reconstitute the lyophilized protein in sterile deionized water or PBS (pH 7.4) to your desired working concentration. Gently swirl to mix — do not vortex — and allow 5 minutes for complete dissolution. Always consult the lot-specific datasheet for the recommended reconstitution volume.

What are the recommended storage conditions for this recombinant protein?

Store the lyophilized protein at –20°C or –80°C upon receipt. After reconstitution, store short-term aliquots at 4°C for up to one week, or prepare single-use aliquots and store at –80°C for long-term use. Avoid repeated freeze–thaw cycles, as these degrade protein activity and integrity.

What experimental applications is Recombinant Human GORAB Protein validated for?

Recombinant Human GORAB Protein (rp373957) is suitable for protein–protein interaction studies (e.g., pull-down, SPR), Western blot as a positive control or standard, ELISA development, functional cell-free assays, and antibody production and validation. Specific validation data are provided on the product datasheet and Certificate of Analysis.

Is Recombinant Human GORAB Protein suitable for clinical or diagnostic use?

No. This product is designated for research use only (RUO) and is not intended or approved for diagnostic, therapeutic, or clinical applications in humans or animals. Researchers must comply with their institutional biosafety and ethics guidelines when using this reagent.

References

1. Hennies HC, et al. Gerodermia osteodysplastica is caused by mutations in SCYL1BP1, a Rab-6 interacting golgin. Nature Genetics. 2008.
2. Witkos TM, Lowe M. The Golgin Family of Proteins. Biochemistry. 2015.

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