Description
Description: Glutamate-rich WD repeat-containing protein 1 (GRWD1) consists of 446 amino acids with molecular mass of 49.4 kDa. It has been found as a protein which interacts with METTL18 and CDT1 proteins. It has been implicated in regulation of DNA replication and/or in ribosome biogenesis.
Applications
- Western blotting (1/1,000~1/3,000 dilution)
- Immunoprecipitation (Assay dependent)
- Immunofluorescence staining / Immunochemistry (1/100~1/1,000 dilution)
Specification
Immunogen: Purified GST-GRWD1 (human, full-length) expressed in E. coli
Purification: The antiserum was first adsorbed with GST-agarose column and then the pass-through fraction was affiinity-purified with GST-GRWD1 agarose column.
Product: 1 mg/ml in PBS and 50% glycerol. Filter-sterilized. Carrier protein and azide free.
Reactivity: Human, mouse and rat. Other species have not been tested.
Storage: Shipped at 4°C. Upon arrival, spin-down and store at -20°C.
Data Link: uniprot/Q9BQ67 (GRWD1_HUMAN); uniprot/Q810D6 (GRWD1_MOUSE)
Reference: This product has been described and used in the following publication:
Sugimoto N. et al. (2008) Identification of novel human Cdt1-binding proteins by a proteomics approach: proteolytic regulation by APC/CCdh1. Mol Biol Cell. 19(3):1007-21.
Fig.1. Identication of GRWD1 proteins in whole cell lysates by western blotting with anti-GRWD1 antibody
Whole cell lysates of HeLa cells untreated (-) and treated (+) with DNA damaging agent, adriamycin (Ad), and NIH3T3 cells were analyzed by Western blotting with anti-GRWD1 antibody at 1/1,000 dilution. The samples were 10 μg. Second antibody was HRP-conjugated goat anti-rabbit IgG used at 1/5,000 dilution. The revelation of multiple bands indicates post-translational modification such as phosphorylation. The level of GRWD1 in the cell was not affected by DNA-damaging treatment. The identity of an additional band at ~85 kDa position other than the GRWD1 band in NIH-3T3 cell lysate is not known. The GRWD1 proteins were identified at a position higher (~55 kDa) than expected from the molecular mass of GRWD1 indicated from cDNA sequence (49.4 kDa).
Fig.2. Immunofluorescence staining of GRWD1 protein in HeLa cells
Hela cells were fixed in 4% paraformaldehyde overnight and permeabilized in 0.25% TritonX 100 in PBS for 10 min. Anti-GRWD1 antibody was used at 1/1,000 dilution. As second antibody, goat anti-rabbit IgG conjugated with Alex488 was used at 1/5,000 dilution. As a signal enhancer, Can Get Signal Immunostain B (Toyobo, Osaka) was used according to the protocol of the supplier. Nuclei were stained with DAPI. GRWD1 protein is localized in nuclei.
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