Recombinant Arabidopsis thaliana Heat shock protein 21 (HSP21) is produced in an E. coli expression system, spanning the full length of the mature protein from amino acids 44 to 227. The protein includes an N-terminal 10xHis-tag and a C-terminal Myc-tag, which streamline purification and detection processes. SDS-PAGE analysis confirms purity greater than 90%, suggesting this preparation should deliver reliable results for research applications.
Heat shock protein 21 (HSP21) in Arabidopsis thaliana appears to be a chloroplast-localized protein that may play a crucial role in how plants respond to heat stress. This protein belongs to the small heat shock protein family, which works to stabilize and refold denatured proteins, helping maintain cellular balance. Research into HSP21 could be vital for understanding plant stress responses and how they adapt to challenging conditions.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
1. In Vitro Protein-Protein Interaction Studies
This dual-tagged HSP21 protein can help researchers investigate how molecular chaperones interact with client proteins or co-chaperones in plant stress response pathways. The N-terminal His-tag makes it possible to immobilize the protein on nickel-affinity surfaces for pull-down assays. Meanwhile, the C-terminal Myc-tag allows detection and measurement using anti-Myc antibodies. Scientists can examine HSP21's binding partners under different temperature conditions to better understand how chloroplasts respond to stress. With purity exceeding 90%, this preparation should give reliable results in co-immunoprecipitation and surface plasmon resonance experiments.
2. Antibody Development and Validation
The recombinant HSP21 protein works well as an antigen for creating specific antibodies against plant heat shock proteins. Its dual-tag system makes purification straightforward and enables quality control during antibody production. Scientists can test antibody specificity using this protein in Western blotting, ELISA, and immunofluorescence applications. Since the mature protein region (44-227aa) represents the functional domain, it's likely suitable for producing antibodies that recognize native HSP21 in actual plant tissues.
3. Biochemical Characterization of Heat Shock Response
This recombinant protein opens doors for detailed biochemical analysis of HSP21 properties. These might include thermal stability, oligomerization states, and structural changes under stress conditions. The His-tag makes protein purification easier for biophysical studies such as dynamic light scattering, circular dichroism spectroscopy, and analytical ultracentrifugation. Researchers can study how the protein behaves across different temperature ranges to understand its role in chloroplastic thermotolerance. The high purity level appears well-suited for quantitative biochemical assays and kinetic studies.
4. Comparative Plant Stress Biology Research
The recombinant Arabidopsis HSP21 may serve as a reference standard for comparing heat shock proteins across different plant species. Scientists can use this protein in cross-reactivity studies to identify conserved regions and functional domains among plant HSP21 orthologs. The dual-tag system provides standardized detection methods across multiple experimental platforms. This application could support evolutionary biology research and help develop universal plant stress markers.
5. Chloroplast Proteomics and Pathway Analysis
This HSP21 protein can work as both a positive control and reference standard in chloroplastic proteomics experiments that investigate heat stress responses. The Myc-tag enables specific detection in complex protein mixtures, while the His-tag allows for controlled spiking experiments in mass spectrometry workflows. Researchers can use this protein to validate proteomic identification methods and measure HSP21 expression levels in plant stress studies. The mature protein sequence should represent the native chloroplastic form in experimental systems.
