|Detection of Mouse RNF168 by Western Blot. Western blot shows lysates of mouse brain (cortex) tissue and mouse brain (thalamus/hypothalamus) tissue. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Mouse RNF168 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7217) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for RNF168 at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|RNF168 in HeLa Human Cell Line. RNF168 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Mouse RNF168 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7217) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
RNF168 (RING [really interesting new gene] finger protein 168; also E3 ubiquitin-protein ligase RNF168) is a 65 kDa (predicted) member of the RNF168 family of proteins. It is ubiquitously expressed, and serves as an E3 ubiquitin ligase. Following DNA damage, RNF8 is recruited to DNA double-strand breaks by phosphoMDC1. Here, RNF8 first monoubiquitinates histone H2A, and then promotes RNF168 recruitment. RNF168 now acts as an additional ubiquitinase, promoting multiple ubiquitinations plus the recruitment of 53BP1, a scaffold protein that holds DNA damage response elements. Mouse RNF168 is 565 amino acids (aa) in length. It contains one Zn-finger/RING domain (aa 16-55) plus two ubiquitin-interacting MIU motifs (aa 168-191 and 438-461). There are three potential Ser/Thr phosphorylation sites. Two potential isoform variants are reported. One shows an alternative start site at Met20, while a second possesses a two aa extension at the N-terminus coupled to a 13 aa substitution for aa 343-565. Over aa 423-565, mouse RNF168 shares 79% and 65.5% aa sequence identity with rat and human RNF168, respectively. Over aa 423-466, mouse RNF168 shares 98% aa sequence identity with human RNF168.
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