Recombinant Human GADD45A Protein: Research Guide & Protocol

Recombinant Human GADD45A Protein is a stress-inducible, DNA damage-response protein produced recombinantly for use in a wide range of in vitro biochemical, cell biology, and signal transduction research applications.

Product Name

Recombinant Human GADD45A Protein

SKU

rp377527

Protein Origin

Human (Homo sapiens)

Protein Class

Stress-inducible / DNA damage response protein

Primary Research Area

DNA damage response, cell cycle regulation, apoptosis, epigenetics

Format

Recombinant protein (lyophilized or liquid; confirm with datasheet)

Biological Background of GADD45A

Growth Arrest and DNA Damage-inducible protein 45 Alpha (GADD45A) is a member of the GADD45 family of small, acidic nuclear proteins. It is encoded by the GADD45A gene and is rapidly induced in response to genotoxic stress, UV radiation, ionizing radiation, hypoxia, and other cellular stressors. GADD45A plays a central role in coordinating several critical cellular responses including cell cycle arrest at the G2/M checkpoint, nucleotide excision repair, and the regulation of apoptosis.

GADD45A interacts with a number of key proteins, including PCNA (Proliferating Cell Nuclear Antigen), p21/CDKN1A, CDK1/CDC2, and MEKK4/MTK1, thereby modulating both DNA repair machinery and stress-activated signaling cascades such as the p38 MAPK and JNK pathways. Additionally, GADD45A has been implicated in active DNA demethylation through its involvement with the base excision repair pathway, linking it to epigenetic regulation.

Due to its multifaceted roles in genome stability and stress signaling, GADD45A is frequently studied in the context of cancer biology, aging, immune regulation, and neurodegenerative disease research.

Reconstitution and Storage Protocol

Proper reconstitution and storage of Recombinant Human GADD45A Protein (rp377527) are essential to preserve biological activity and ensure experimental reproducibility. The following guidelines reflect standard best practices for recombinant proteins; always refer to the product-specific datasheet supplied with each lot for definitive instructions.

• Reconstitution: If supplied in lyophilized form, briefly centrifuge the vial before opening to collect the powder. Reconstitute in sterile, ultrapure water or an appropriate buffer (e.g., phosphate-buffered saline, pH 7.4, with a carrier protein such as 0.1% BSA if needed for stability). Gently pipette to mix; avoid vortexing to prevent protein denaturation.
• Working Concentration: Prepare aliquots at concentrations appropriate for your assay. Consult the datasheet for recommended concentration ranges.
• Short-term Storage: Store reconstituted protein at 4°C for up to 7 days.
• Long-term Storage: For extended storage, aliquot and freeze at −20°C or −80°C. Avoid repeated freeze-thaw cycles, which can significantly reduce activity.
• Carrier Protein: Adding 0.1–0.5% BSA or human serum albumin to the reconstitution buffer can improve protein stability during storage.

Note: Specific buffer composition, concentration, and storage conditions must be confirmed from the official Aladdin Scientific product datasheet for lot rp377527.

Experimental Applications

Recombinant Human GADD45A Protein (rp377527) is suitable for multiple research applications leveraging its role in DNA damage signaling and stress response pathways:

• Biochemical Assays: Use as a standard or positive control in ELISA, Western blotting, or dot blot assays to validate antibody specificity against GADD45A.
• Protein–Protein Interaction Studies: Employ in co-immunoprecipitation (Co-IP), pull-down assays, or surface plasmon resonance (SPR) experiments to characterize interactions with binding partners such as PCNA, p21, or CDK1.
• Functional/Activity Assays: Investigate GADD45A's role in modulating kinase activity (e.g., CDK1 inhibition) or nucleotide excision repair in cell-free systems.
• Cell-Free Transcription/Translation Systems: Serve as a reference protein in in vitro transcription or translation experiments studying stress-response gene networks.
• Structural Studies: Suitable as a starting material for biophysical characterization including circular dichroism (CD) spectroscopy or dynamic light scattering (DLS).
• Antibody Production & Validation: Use as an immunogen or coating antigen for generating and validating GADD45A-specific antibodies.

Researchers should optimize protein concentrations and assay conditions empirically for each specific application.

Quality, Purity & Regulatory Considerations

Aladdin Scientific's Recombinant Human GADD45A Protein (rp377527) is manufactured to research-grade quality standards. Key quality attributes typically assessed for recombinant proteins of this class include:

• Purity: Typically assessed by SDS-PAGE and/or HPLC. Target purity for research-grade recombinant proteins is generally ≥85–95%; consult the Certificate of Analysis (CoA) for lot-specific data.
• Identity Confirmation: Verified by Western blot or mass spectrometry against the expected molecular weight of human GADD45A (approximately 18–21 kDa, depending on expression system and any tags).
• Endotoxin Level: Endotoxin content should be assessed (typically by LAL assay) and confirmed on the CoA, particularly important for cell-based assays.
• Biological Activity: Functional validation data, where available, should be reviewed on the product datasheet.
• Research Use Only (RUO): This product is intended for research purposes only and is not suitable for diagnostic, therapeutic, or clinical use.

Laboratory managers should request a current CoA and Safety Data Sheet (SDS) at the time of ordering. Storage, handling, and disposal must comply with applicable institutional and local safety regulations.

Frequently asked questions

What is GADD45A and why is it important in research?

GADD45A (Growth Arrest and DNA Damage-inducible protein 45 Alpha) is a stress-response nuclear protein that plays key roles in DNA repair, cell cycle checkpoint activation (particularly at G2/M), apoptosis regulation, and epigenetic control via DNA demethylation. It is widely studied in cancer biology, genotoxicity, immune signaling, and aging research, making recombinant GADD45A protein a valuable tool for mechanistic in vitro studies.

How should I reconstitute and store Recombinant Human GADD45A Protein (rp377527)?

Reconstitute lyophilized protein in sterile ultrapure water or an appropriate buffer such as PBS (pH 7.4), gently mixing without vortexing. Aliquot and store long-term at −80°C to avoid repeated freeze-thaw cycles. For short-term use, the protein can be kept at 4°C for up to 7 days. Always consult the product-specific datasheet and CoA for definitive reconstitution and storage instructions for your specific lot.

What applications is Recombinant Human GADD45A Protein suitable for?

Recombinant Human GADD45A Protein (rp377527) is suitable for Western blot controls, ELISA development, protein–protein interaction studies (Co-IP, pull-down, SPR), functional cell-free assays, antibody production and validation, and biophysical characterization. Researchers should optimize conditions for each specific assay type.

Is Recombinant Human GADD45A Protein suitable for clinical or diagnostic use?

No. Recombinant Human GADD45A Protein (rp377527) from Aladdin Scientific is classified as Research Use Only (RUO). It is not approved, validated, or intended for diagnostic, therapeutic, or clinical applications. Use must comply with your institution's research and safety regulations.

Where can I find lot-specific quality data for rp377527?

Lot-specific quality data, including purity by SDS-PAGE, identity confirmation, endotoxin levels, and biological activity (where available), are provided in the Certificate of Analysis (CoA) supplied with each shipment. You may also request a CoA from Aladdin Scientific's customer service team at the time of ordering or after receipt.

References

1. Fornace AJ Jr, et al. Mammalian genes coordinately regulated by growth arrest signals and DNA-damaging agents. Molecular and Cellular Biology. 1989;9(10):4196–4203.
2. Hollander MC, et al. Genomic instability in Gadd45a-deficient mice. Nature Genetics. 1999;23(2):176–184.
3. Barreto G, et al. Gadd45a promotes epigenetic gene activation by repair-mediated DNA demethylation. Nature. 2007;445(7128):671–675.
4. Zhan Q, et al. Association with Cdc2 and inhibition of Cdc2/Cyclin B1 kinase activity by the p53-regulated protein Gadd45. Oncogene. 1999;18(18):2892–2900.

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