Bethyl Laboratories, Inc.

Antibodies to the
WD-repeat Family of Proteins

March 25,2010

Overview

The WD-repeat family of proteins is a large family of proteins whose members are structurally homologous but functionally diverse. The WD-repeat motif is defined by four or more repeating units of a conserved core of 40-60 amino acids that begins with a glycine-histidine (GH) dipeptide and ends with a tryptophan-aspartic acid (WD) dipeptide. Regions within the WD-repeat exhibit sequence heterogeneity and are likely responsible for imparting their functional diversity. WD-repeat-containing proteins perform in a spectrum of cellular activities which includes RNA processing, signal transduction, transcription, vesicular trafficking, cytoskeletal assembly, chromatin assembly, and various aspects of cell division.

The crystal structure of the beta subunit of the heterotrimeric G-protein has been resolved and has provided a structural characterization of the WD-repeat. Each of the seven repeats of the G-protein beta subunit folds into small anti-parallel beta-sheets that radiate from a central pseudo-symmetrical axis to form a beta propeller structure. The surfaces formed by the beta propeller structure are proposed to function as a scaffold capable of coordinating reversible protein-protein interactions.

Protein-protein interactions provide a source of complexity for the multitude of physiological processes that must be executed by eukaryotes for development and for the maintenance of homeostasis. Through their conserved beta propeller structure, WD-repeat-containing proteins contribute to this source of complexity and have thus established themselves as a critical subject of study.

Bethyl Laboratories' Portfolio
APC4 BOP1 Bub3 CAF-1 p60 CDC20 COP1/RFWD2 Coronin 1 Coronin 2 CSTF50 DMWD DTL/CDT2 EIF3S9/eIF3B EML4 FBW7 GbetaL Gemin5 GRWD1/WDR28 GTF3C2/TFIIIC110	HERC1 LIS-1 LRWD1 NRIP - NEW NUP214 PIP1/WDR84 PLRG1 PPP2R2B PRP19/PSO4 RACK1 - NEW Raptor RbBP4 RbBP5 RBBP7 RCD8 RFWD3 SEC31A - NEW TBLR1	TRIM3/BERP UTP18/WDR50 VprBP WDHD1/AND-1 WDR3 WDR4 WDR5 - NEW WDR26 - NEW WDR33 WDR42A WDR43 - NEW WDR44 WDR46 WDR62 WDR77/MEP50 WDR79 WDR91 ZFP106


Detection of Human BUB3 by IHC. Samples: FFPE section of human metastatic lymph node. Antibodies: Affinity purified rabbit anti-BUB3 (Cat. No. IHC-00123) used at a dilution of 1:250. Detection: DAB staining using Immunohistochemistry Accessory Kit (Cat. No. IHC-101).	Detection of Human and Mouse RACK1 by WB (h&m) and IP (h). Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 mcg), and mouse NIH3T3 (M; 50 mcg) cells. Antibodies: Affinity purified rabbit anti-RACK1 antibody A302-545A used for WB at 0.1 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 3 mcg/mg lysate. RACK1 was also immunoprecipitated by rabbit anti-RACK1 antibody BL9179, which recognizes a downstream epitope. For blotting immunoprecipitated RACK1, ReliaBLOT� Reagents and Procedures (Cat. No. WB120) were used. Detection: Chemiluminescence with exposure times of 3 minutes (A) and 30 seconds (B).	Detection of Human WDR77/MEP50 by lHC-IF. Samples: FFPE section of human prostate carcinoma. Antibodies: Affinity purified rabbit anti-WDR77/MEP50 (Cat. No. IHC-00338) used at a dilution of 1:100. Detection: Red-fluorescent goat anti-rabbit IgG highly cross-adsorbed Antibody Hilyte PlusTM 555 (A120-501E) used at a dilution of 1:100.

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