1. Basic information of recombinant protein SIRPB1A.
SIRPB1A (Signal-Regulatory Protein Beta 1A) is a transmembrane glycoprotein that is widely expressed in a variety of cell types, including immune cells and neurons. SIRPB1A plays an important role in cell-to-cell signaling, and its occurrence and development, axonal guidance of neurons, and regulation of inflammatory responses are closely related. In order to further study the function and mechanism of SIRPB1A, we plan to perform expression and purification experiments for the recombinant protein SIRPB1A.
Catalog No. PA1000-5534
2. Experimental design.
1.Clone the sirpb1a gene.
The sirpb1a gene was amplified from the corresponding DNA template using PCR and digested before being inserted into the appropriate expression vector. Sequencing is performed to confirm that the cloned gene sequence is correct.
2.Transfection and expression.
Transfect the expression vector containing the SIRPB1A gene into a suitable cell line (e.g., HEK293 or CHO cells) and select the appropriate expression conditions (e.g., media composition, temperature, and cell density).
3.Purification of the recombinant protein SIRPB1A
The culture supernatant was collected and the recombinant protein SIRPB1A was purified by affinity chromatography or ion exchange chromatography. Sample purity was detected by SDS-PAGE and Western blot.
4.Functional assessment.
4.1 Cell proliferation assay: MTT or CCK-8 method was used to detect the effect of different concentrations of recombinant protein SIRPB1A on cell proliferation.
4.2 Binding experiments: Binding between a recombinant protein and its potential ligand, such as neuronal growth factor or specific receptor, using methods such as ELISA or surface plasmon resonance (SPR).
4.3 Immunomodulation assays: In vitro or in vivo models, the effects of recombinant protein SIRPB1A on the activation, proliferation, differentiation and corresponding molecular signaling pathways of immune cells were studied.
V. Expected Results.
Through this experiment, we hope to successfully express and purify the functionally active recombinant protein SIRPB1A, and explore its role in cell-to-cell signaling, neuronal axon guidance, disease occurrence and **. At the same time, we also hope to unravel the functional mechanism of SIRPB1A in immune cell activation and regulation of inflammatory responses.
In summary, a comprehensive set of experimental protocols for the expression, purification, and functional study of SIRPB1A recombinant protein was designed to gain a deeper understanding of the biological characteristics and functions of the protein.