Cat. No.
MABL-2203
Application
IP, WB, IF
Isotype
Engineer antibody
Species Reactivity
Human
Clone No.
Hec1 (9G3)
From
Recombinant Antibody
Specificity
This antibody specifically binds amino acids 200–215 in the N terminal globular domain of the human Hec1 protein, exterior to Spc24 at metaphase kinetochores. It binds in a region which resides within the well-ordered calponin homology domain of the protein. Hec 1 protein acts as a component of the essential kinetochore-associated NDC80 complex, which is required for chromosome segregation and spindle checkpoint activity. HEC protein is expressed most abundantly in the S and M phases of rapidly dividing cells but not in terminal differentiated cells. It localizes to the nuclei of interphase cells, and a portion distributes to centromeres during M phase. Inactivation of HEC by microinjection of specific monoclonal antibodies into cells during interphase severely disturbs the subsequent mitoses.
Alternative Names
NDC80; HEC; HEC1; KNTC2; Highly expressed in cancer protein; Kinetochore protein Hec1; HsHec1; Kinetochore-associated protein 2; Retinoblastoma-associated protein HEC
UniProt
O14777
Immunogen
The original antibody was generated in mice by immunization with a purified protein fragment encompassing amino acids 56–632 of the human Hec1 protein.
Application Notes
This antibody is widely used in studies involving the role of Hec1 protein in mitosis. This antibody can immunoprecipitate human Hec1 protein from nuclear, cytoplasmic, membrane and other components of T24 human bladder carcinoma cells. This antibody was capable of recognizing a 76-KDa protein in western blot. It was reported that when T24 human bladder carcinoma cells were synchronized in S phase and microinjected with this antibody the cells contained multiple, fragmented nuclei (PMID: 9315664). Preincubation of the antibody with C-2 peptide (comprising amino acids 200-215 of Hec1) prevented the antibody from recognizing kinetochores in immunofluorescence assays. Immunofluorescent staining of HeLa and PtK1 cells in metaphase with 9G3 and another anti-Spc24 antibody revealed that the Nterminus of Hec1 was always exterior to Spc24 at kinetochores of bi-oriented chromosomes. It was further reported that PtK1 cells injected with this antibody exhibited mitotic defects (PMID: 17129782). A recombinantly generated version of this antibody rMAb-Hec1 robustly recognized kinetochores however, the original antibody exhibited more spindle staining and overall background staining when compared to the recombinant version (PMID: 34970967).
Antibody First Published
Chen et al. HEC, a novel nuclear protein rich in leucine heptad repeats specifically involved in mitosis. Mol Cell Biol. 1997 Oct; 17(10): 6049–6056. PMID:9315664
Note on publication
Describes the use of this antibody to study the importance of Hec1 protein in mitosis.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
