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Mad1 Mouse mAb

Catalog No.: F3921

Application: Reactivity: human

Usage Information

Dilution
WB1:2000

Application
WB, IF

Reactivity
human

Source
Mouse

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
~83 kDa

Datasheet & SDS

Biological Description

Specificity
Mad1 Mouse mAb detects endogenous levels of total Mad1 protein.

Clone
M18F4

Synonym(s)
Mitotic spindle assembly checkpoint protein MAD1, Mitotic arrest deficient 1-like protein 1, MAD1-like protein 1, Mitotic checkpoint MAD1 protein homolog, HsMAD1, hMAD1, Tax-binding protein 181

Background
Mad1 (Mitotic Arrest Deficient 1) is a crucial component of the spindle assembly checkpoint (SAC) family, ensuring precise chromosome segregation by preventing anaphase initiation until all chromosomes are correctly attached to spindle microtubules. Mad1 is a 718-amino acid homodimeric coiled-coil protein, with the C-terminal domain (CTD) playing a pivotal role in its function and regulatory control. The CTD harbors motifs vital for Mad1’s growth-inhibitory activity and ability to repress Myc-dependent transcription. Deletion of its last 18 amino acids (region V) abolishes these functions; however, deleting both region V and adjacent region IV restores them, revealing a complex phosphorylation-regulated interplay within these domains. The CTD also facilitates Mad1’s kinetochore targeting and interaction with checkpoint proteins like Bub1, forming a multivalent interface essential for recruiting Mad2 to unattached kinetochores. This recruitment catalyzes Mad2 conformational activation and mitotic checkpoint complex (MCC) assembly, which inhibits the anaphase-promoting complex/cyclosome (APC/C), thereby delaying anaphase until proper attachment is achieved. Regulatory phosphorylation by kinases such as Mps1 and Bub1 modulates Mad1’s activity, while p31^comet^ serves as an inhibitor once kinetochores are correctly attached. Mad1 also influences transcriptional repression through interaction with Myc-Max complexes, impacting cell cycle progression. Dysregulation of Mad1 functions leads to chromosome instability and aneuploidy, contributing to tumorigenesis.

Background

Mad1 (Mitotic Arrest Deficient 1) is a crucial component of the spindle assembly checkpoint (SAC) family, ensuring precise chromosome segregation by preventing anaphase initiation until all chromosomes are correctly attached to spindle microtubules. Mad1 is a 718-amino acid homodimeric coiled-coil protein, with the C-terminal domain (CTD) playing a pivotal role in its function and regulatory control. The CTD harbors motifs vital for Mad1’s growth-inhibitory activity and ability to repress Myc-dependent transcription. Deletion of its last 18 amino acids (region V) abolishes these functions; however, deleting both region V and adjacent region IV restores them, revealing a complex phosphorylation-regulated interplay within these domains. The CTD also facilitates Mad1’s kinetochore targeting and interaction with checkpoint proteins like Bub1, forming a multivalent interface essential for recruiting Mad2 to unattached kinetochores. This recruitment catalyzes Mad2 conformational activation and mitotic checkpoint complex (MCC) assembly, which inhibits the anaphase-promoting complex/cyclosome (APC/C), thereby delaying anaphase until proper attachment is achieved. Regulatory phosphorylation by kinases such as Mps1 and Bub1 modulates Mad1’s activity, while p31^comet^ serves as an inhibitor once kinetochores are correctly attached. Mad1 also influences transcriptional repression through interaction with Myc-Max complexes, impacting cell cycle progression. Dysregulation of Mad1 functions leads to chromosome instability and aneuploidy, contributing to tumorigenesis.

References
https://pubmed.ncbi.nlm.nih.gov/24477934/ https://pubmed.ncbi.nlm.nih.gov/10825189/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%