MABS2250

Anti-phospho-VCP (Ser784) Antibody, clone 3E4

• Synonym(s): Transitional endoplasmic reticulum ATPase;EC:3.6.4.6;TER ATPase;15S Mg(2+)-ATPase p97 subunit;Valosin-containing protein
• UNSPSC Code: 12352203
• NACRES: NA.43

Properties

• biological source: mouse
• Quality Level: 200
• conjugate: unconjugated
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 3E4, monoclonal
• mol wt: calculated mol wt 89.32 kDa; observed mol wt ~95 kDa
• purified by: using protein G
• species reactivity: human
• species reactivity (predicted by homology): bovine, porcine, canine, rat, monkey, mouse

Description

General description

Transitional endoplasmic reticulum ATPase (UniProt:P55072; also known as EC:3.6.4.6, TER ATPase, 15S Mg(2+)-ATPase p97 subunit, Valosin-containing protein, VCP) is encoded by the VCP gene (Gene ID: 7415) in human. VCP is a homohexameric protein that forms a ring-shaped particle and displays a six-fold radial symmetry. It is shown to be essential for the fragmentation of Golgi stacks during mitosis and for their reassembly after mitosis. It is involved in the formation of the transitional endoplasmic reticulum (tER). The transfer of membranes from the endoplasmic reticulum to the Golgi apparatus occurs via 50-70 nm transition vesicles, which derive from part-rough, part-smooth transitional elements of tER. Vesicle budding from the tER is reported to be an ATP-dependent process. It is shown that the ternary complex containing UFD1, VCP and NPLOC4 regulates spindle disassembly at the end of mitosis. This complex binds ubiquitinated proteins and is necessary for the export of misfolded proteins from the ER to the cytoplasm, where they are degraded by the proteasome. VCP is also involved in DNA damage response. It is recruited to double-strand breaks sites in a RNF8- and RNF168-dependent manner and promotes the recruitment of TP53BP1 at DNA damage sites. It is recruited to stalled replication forks by DNA-binding metalloprotease SPRTN and together they are involved in the repair of covalent DNA-protein cross-links during DNA synthesis. VCP is also required for cytoplasmic retro-translocation of stressed/damaged mitochondrial outer-membrane proteins and their subsequent proteasomal degradation. DNA damage is reported to induce phosphorylation of serine 784 that selectively enhances chromatin-associated protein degradation and is required for DNA repair, signaling, and cell survival. These functional effects of serine 784 phosphorylation correlate with a decrease in VCP association with chromatin, cofactors NPL4/UFD1, and polyubiquitinated substrates. (Ref.: (Ref.: Zhu, C., et al.(2020). Cell Rep. 31(10); 107745; Fielden, J., et al. (2020). Nat. Commun. 11(1); 1274).

Specificity

Clone 3E4 is a mouse monoclonal antibody that detects VCP phosphorylated on serine 784.

Immunogen

KLH-conjugated linear peptide corresponding to 14 amino acids surrounding phosphoserine 784 from the C-terminal region of human Transitional endoplasmic reticulum ATPase (VCP).

Application

Quality Control Testing

Evaluated by Western Blotting in lysates from HeLa cells treated with etoposide.

Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected VCP phosphorylated on serine 784 in lysates from HeLa cells treated with etoposide (10 μM; 24 h), but not in untreated cells.

Tested Applications

Western Blotting Analysis: A representative lot detected phospho-VCP (Ser784) in Western Blotting applications (Zhu, C., et al. (2020). Cell Rep. 31(10):107745).

Immunoprecipitation Analysis: A representative lot immunoprecipitated phospho-VCP (Ser784) in Immunoprecipitation applications (Zhu, C., et al. (2020). Cell Rep. 31(10):107745).

Immunocytochemistry Analysis: A representative lot detected phospho-VCP (Ser784) in Immunocytochemistry applications (Zhu, C., et al. (2020). Cell Rep. 31(10):107745).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-phospho-VCP (Ser784), clone 3E4, Cat. No. MABS2250, is a mouse monoclonal antibody that detects Valosin-containing protein (VCP) phosphorylated at serine 784 and is used in Immunocytochemistry, Immunoprecipitation, and Western Blotting.

Physical form

Purified mouse monoclonal antibody IgG2b in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable