Anti-RAG1 Rabbit Polyclonal Antibody for WB, ELISA - P15918

Core Product Specifications and Parameters

Biological Background: RAG1 Function and Localization

• RAG1 (Recombination activating protein 1, also known as RAG-1 and RNF74) is the catalytic component of the RAG complex, essential for V(D)J recombination that assembles diverse immunoglobulin and T-cell receptor genes in developing lymphocytes.
• It directly binds to conserved recombination signal sequences (RSS) and catalyzes site-specific DNA double-strand breaks through a two-step mechanism: initial nicking upstream of the heptamer, followed by hairpin formation via transesterification, creating blunt signal ends and hairpinned coding ends. RAG2 is required for all catalytic activities.
• Beyond endonuclease activity, RAG1 functions as an E3 ubiquitin-protein ligase that monoubiquitinates histone H3 at unknown sites, a modification required for the joining step of V(D)J recombination. Additionally, trimethylation of H3 at Lys-4 (H3K4me3) stimulates both cleavage steps.
• RAG1 localizes to the nucleus, where it interacts with chromatin to mediate recombination events.
• Its expression is restricted to maturing lymphoid cells, consistent with its role in lymphocyte development.
• RAG1 also participates in pre-B cell allelic exclusion by inducing ATM-dependent repositioning of one immunoglobulin allele to pericentromeric heterochromatin, preventing recombination of the second allele and ensuring clonality.
• The protein undergoes alternative splicing, and its multifunctional nature is underscored by keywords such as DNA-binding, endonuclease, hydrolase, metal-binding (zinc-finger), transferase, ubl conjugation, and disease variants linked to severe combined immunodeficiency (SCID).

Experimental Guidance and Technical Tips

• The immunogen spans the N-terminal region (amino acids 1-270) of human RAG1, which includes the DNA-binding domain. This region may detect multiple isoforms arising from alternative splicing.
• For Western blotting, the predicted molecular weight is ~119 kDa. Consider using a 6-8% SDS-PAGE gel and extended transfer times for high-molecular-weight proteins.
• ELISA protocols may utilize recombinant RAG1 protein or peptide coating; validate the assay conditions in your sample matrix to ensure specificity.
• Cross-reactivity with mouse is expected; validate performance in mouse lymphoid tissues or cell lines if used in these systems.
• Protein extracts from human or mouse lymphoid cell lines (e.g., Jurkat, Raji, EL4) are likely suitable positive controls; non-lymphoid cell lines can serve as negative controls.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers, laboratories, and research institutes with one-stop access to IVD raw materials, technical services, and consulting—covering every stage from concept to clinic. Focusing on RAG1 and immune recombination targets, we offer validated antibody pairs, optimized polyclonal and monoclonal antibodies, bulk ancillary reagents, and rare target raw-material sourcing to support your V(D)J recombination, immunodeficiency disorder, and lymphocyte development research and assay development. Let CamelBio bridge your sourcing needs efficiently.