Monoclonal Antibodies: ASF1a, HEF1/NEDD9/Cas-L, hBUBs

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Monoclonal Antibodies against human ASF1a, a histone chaperone and regulator of cell proliferation

ASF1a is a human ortholog of the yeast Asf1p protein, a histone chaperone and a binding partner of HIRA. Like HIRA, ASF1a contributes to formation of heterochromatin in senescent human cells, and also has roles in DNA replication, repair and transcription.

ClonesMPH7, MPB1, MPD5
Applications:Each clone works for IP. None works for WB
Specificity:Each reacts with human protein. Reactivity against other species has not yet been fully characterized
Immunogen:Glutathione S-transferase-ASF1a (human ASF1a)

Clone MPH7: IgG2b

Clones MPB1 and MPD5: IgG1

Citation:Zhang R, Poustovoitov MV, et al. "Formation of MacroH2A-containing senescence-associated heterochromatin foci and senescence driven by ASF1a and HIRA," Dev Cell. 2005 Jan;8(1):19-30 [PMID: 15621527]

HEF1/NEDD9/Cas-L, A Novel Signal Transduction Molecule Associated with Cell Motility, Growth, Apoptosis and Oncogenic Transformation



Dr. Erica Golemis and colleagues have discovered and characterized HEF1/NEDD9/Cas-L (Human Enhancer of Filamentation 1/Neural precursor cell expressed, developmentally downregulated 9), a member of the Cas family of docking proteins that also includes p130Cas and Efs/Sin. HEF-1 is a multifunctional protein involved in integrin-based signaling that affects cell motility, growth, apoptosis and oncogenic transformation. Experiments convey the idea that HEF1 may represent a marker for an enhanced possibility of tumor metastasis. Furthermore, HEF1 belongs to the 18 gene signature prognostic of breast cancer metastasis to lung tissue.
U.S. patents have been granted that broadly cover HEF-1. Fox Chase Cancer Center is offering licenses to HEF-1 cell lines, antibodies, reagents and patents for both research and commercial use.

Applications:WB, IF, and IP
Specificity:Specific only for HEF1, does not recognize p130Cas. Have not tested on Sin1. Tested only on human (MCF7, Hela) and mouse cell lines (MEF).

IgG1 kappa

Notes:All applications were tested on human and mice cell lines. IF was performed on 4%PFA fixed cells or methanol. This mAb mostly detects HEF1 localized at the focal adhesion sites.
Citation:Pugacheva EN, and Golemis EA, "The focal adhesion scaffolding protein HEF1 regulates activation of the Aurora-A and Nek2 kinases at the centrosome," Nat. Cell Biol 2005 Sep; 7, 937 - 946 [PMID: 16184168]

Antibodies against Human Kinase Proteins hBUB1A, hBUB1B, and hBUBR1

The BUB family of serine/threonine kinases is a major target of oncology therapeutic research and development. hBUB1 and hBUBR1 are kinases that associate with hBUB3 and are essential components of the mitotic checkpoint. hBUBR1 appears to provide two checkpoint functions by monitoring CENP-E-dependent activities at the kinetochore and by regulating cyclosome/APC activity.

hBUBR1 is required in order for microtubule inhibitors to arrest cells in mitosis. Non-mutational inactivation of hBUB1 and hBUBR1 appear related to stage of colon cancer and could be related to aneuploidy in human colorectal and other cancers.

Applications:WB, IP and IF
Citation:Chan GK, Jablonski SA, et al. "Human BUBR1 is a mitotic checkpoint kinase that monitors CENP-E functions at kinetochores and binds the cyclosome/APC," J Cell Biol Sept 1999;146(5):941-54 [PMID: 10477750]