ABS1583

Anti-VprBP Antibody

from rabbit, purified by affinity chromatography

• Synonym(s): Protein VPRBP, DDB1- and CUL4-associated factor 1, HIV-1 Vpr-binding protein, Serine/threonine-protein kinase VPRBP, Vpr-interacting protein, VprBP
• UNSPSC Code: 12352203
• eCl@ss: 32160702

Properties

• biological source: rabbit
• antibody form: affinity isolated antibody
• antibody product type: primary antibodies
• clone: polyclonal
• purified by: affinity chromatography
• species reactivity: human, mouse
• technique(s): immunoprecipitation (IP): suitable; western blot: suitable
• NCBI accession no.: NP_055518
• UniProt accession no.: Q9Y4B6
• shipped in: ambient
• target post-translational modification: unmodified
• Gene Information: human ... VPRBP(9730)

Description

General description

Protein VPRBP (EC 2.7.11.1; UniProt Q9Y4B6; also known as DDB1- and CUL4-associated factor 1, HIV-1 Vpr-binding protein, Serine/threonine-protein kinase VPRBP, Vpr-interacting protein, VprBP) is encoded by the VPRBP (also known as DCAF1, KIAA0800, RIP) gene (Gene ID 9730) in human. VprBP/DCAF1 acts as an adaptor molecule that mediates substrates recruitment for both the RING-type CRL4 (Cul4-Roc1-DDB1) and HECT-type EDD/UBR5 E3 ubiquitin ligase complexes. CRL4 substrates known to be recruited by VprBP include Merlin, Mcm10, UNG2, SMUG1, RORαme1. Known EDD/UBR5 substrates recruited by VprBP include Katanin and TERT. VprBP is involved in a variety of normal cellular processes, including proliferation, DNA replication, cell cycle progression, telomere maintenance, and DNA damage responses. All three TET methylcytosine dioxygenases are targeted by CRL4-VprBP for monouquitination on a highly conserved lysine residue (human TET1 K1589, TET2 K1299, and TET3 K859), an essential event for TET chromatin binding, Vprbp gene knockout in mouse oocytes abrogates paternal DNA hydroxymethylation in zygotes, and the monoubiquitylation site is among multiple recurrent TET2-inactivating mutations targeted by leukemia. As indicated by its name, VprBP is also a well known target utilized by the Vpr and Vpx accessory factors of primate lentiviruses to highjack the host CRL4-VprBP E3 ligase function.

Specificity

Specificity was verified by reduced target band detection upon shRNA-mediated VprBP knockdown in U2OS, WI-38/E6, HeLa, and HEK293T cells, as well as following cre expression in MEFs harboring floxed Vprbp alleles (McCall, C.M., et al. (2008). Mol. Cell. Biol. 28(18):5621-5633; Nakagawa, T., et al. (2015). 57(2):247-260). Target region is present in human spliced isoforms 1-4 as well as mouse isoforms 1 and 2 reported by UniProt (Q9Y4B6, Q80TR8), but missing in mouse isoforms 3 and 4 and not present in rat VprBP (NM_001191798.1 & NP_001178727.1).

Immunogen

Epitope: C-terminus

KLH-conjugated linear peptide corresponding to human VprBP C-terminal end sequence.

Application

Detect Protein VprBP using this rabbit polyclonal Anti-VprBP Antibody, Cat. No. ABS1583, validated for use in Immunoprecipitation and Western Blotting.

Immunoprecipitation Analysis: A representative lot co-immunoprecipitated TET1 and TET2 with VprBP from human monocyte, mouse embryonic fibroblast and stem cell (MEF and mESC) lysates (Nakagawa, T., et al. (2015). 57(2):247-260).

Immunoprecipitation Analysis: A representative lot co-immunoprecipitated DDB1 and NEDD8-modified CUL4A with VprBP from lysate of proteasome inhibitor MG132-treated U2OS human osteosarcoma cells (McCall, C.M., et al. (2008). Mol. Cell. Biol. 28(18):5621-5633).

Western Blotting Analysis: A representative lot detected endogenous VprBP co-immunoprecipitated with FLAG-tagged full-length as well as C-terminal cysteine-rich dioxygenase (CD) domain constructs of murine TET1, TET2, and TET3 exogenously expressed in HEK293T transfectants. Target band detection was abolished in lysates from VprBP shRNA-treated HEK293T cells (Nakagawa, T., et al. (2015). 57(2):247-260).

Western Blotting Analysis: A representative lot detected VprBP in CUL4A immunoprecipitates from human breast cancer BT474 and osteosarcoma U2OS cell lysates. Endogenous VprBP co-immunoprecipitated only with Myc-tagged CUL4A, but not CUL1/2/3/5, exogenously expressed in HEK293T transfectants (McCall, C.M., et al. (2008). Mol. Cell. Biol. 28(18):5621-5633).

Western Blotting Analysis: A representative lot detected shRNA-mediated VprBP knockdown in U2OS, WI-38/E6, and HeLa cells. Cellular DDB1 knockwon resulted in a 72% and 58% drop of VprBP level in HCT116 whole cell lysate and chromatin preparation, respectively (McCall, C.M., et al. (2008). Mol. Cell. Biol. 28(18):5621-5633).

Western Blotting Analysis: A representative lot detected VprBP knockdown upon retrovirus-mediated cre expression in mouse embryonic fibroblasts (MEFs) harboring floxed Vprbp alleles (McCall, C.M., et al. (2008). Mol. Cell. Biol. 28(18):5621-5633).

Research Category
Signaling

Quality

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected VprBP in 10 µg of HeLa cell lysate.

Target description

~169 kDa observed. 169.0/168.9/118.2 kDa (human isoform 1/2/3) and 168.9/169.7 kDa (mouse isoform 1/2) calculated. Uncharacterized bands may be observed in some lysate(s).

Physical form

Affinity purified.

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

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

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: 12 - Non Combustible Liquids
• wgk_germany: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable