Anti-MAD2/MAD2L1 Polyclonal Antibody for IHC-P, IF/ICC, ELISA - Q13257

Core Product Specifications and Parameters

Biological Background: MAD2/MAD2L1 Function and Localization

• MAD2L1 encodes mitotic spindle assembly checkpoint protein MAD2A (also known as MAD2-like protein 1), a critical regulator of chromosome segregation fidelity.
• MAD2L1 is a core component of the mitotic checkpoint complex (MCC), which inhibits the anaphase-promoting complex/cyclosome (APC/C) to delay chromosome segregation until all chromosomes are correctly attached to the mitotic spindle. Related references: PMID:10700282 PMID:11804586 PMID:15024386
• The MCC, consisting of CDC20, MAD2L1/MAD2, BUB1B/BUBR1, and BUB3, serves as the key effector of the spindle assembly checkpoint. Related references: PMID:15020684 PMID:27509861
• MCC-bound APC/C remains inactive, preventing the metaphase-to-anaphase transition; checkpoint silencing triggers UBCH10-mediated auto-ubiquitination of CDC20, leading to APC/C reactivation. Related references: PMID:27509861
• MAD2L1 undergoes a dynamic conformational switch: the closed conformer (C-MAD2) forms a complex with MAD1L1 at unattached kinetochores, recruiting open MAD2 (O-MAD2) and catalyzing its conversion to C-MAD2, thereby amplifying checkpoint signaling. Related references: PMID:29162720
• Subcellularly, MAD2L1 localizes to the nucleus, kinetochores at centromeres, spindle poles, and the cytoplasm, reflecting its multifaceted mitotic functions.
• The protein is subject to acetylation and phosphorylation, and alternative splicing generates multiple isoforms; these modifications and variants may modulate checkpoint activity and protein interactions.

Experimental Guidance and Technical Tips

• The immunogen corresponds to the N-terminal 205 amino acids of human MAD2L1; consider verifying isoform and species specificity in your particular assay system, as cross-reactivity to closely related proteins may occur.
• For immunohistochemistry (IHC-P), antigen retrieval methods and fixation conditions can influence staining; optimization is recommended using appropriate positive (e.g., mitotic tissues) and negative controls.
• In immunofluorescence (IF/ICC), expect kinetochore and spindle pole labeling during mitosis; co-staining with centromere or tubulin markers can help confirm target engagement.
• For ELISA, purified recombinant MAD2L1 protein can serve as a positive control; titration of antibody and antigen concentrations is advised to establish optimal signal-to-noise ratios.
• Always validate the antibody in your specific samples and application context, as performance may vary across cell types and experimental conditions.

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