Analysis Note

ControlHeLa cell lysate

Application

Use Anti-RNF168 Antibody (Rabbit Polyclonal Antibody) validated in WB, ICC, IF to detect RNF168 also known as E3 ubiquitin-protein ligase RNF168.

Research CategoryEpigenetics & Nuclear Function

Research Sub CategoryChromatin Biology

Immunocytochemistry Analysis: 1.0 µg/mL from a representative lot detected RNF168 in HeLa, A431, HUVEC, and NIH/3T3 cells.
Immunohistochemistry Analysis: A representative lot detected RNF168 in frozen human lung adenocarcinoma tissue sections (Xie, X., et al. (2018) Nat. Cell Biol. 20(3); 320-331).

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.

General description

E3 ubiquitin-protein ligase RNF168 (UniProt Q8IYW5; also known as hRNF168, RING finger protein 168) is encoded by the RNF168 gene (Gene ID 165918) in human. Ubiquitylation/Ubiquitination is an essential early signal in the DNA damage repair process. Multiple E3 ligases, including RNF2 (RING2), RNF8 and RNF168, mediate the addition of K63-linked polyubiquitin chains to histone 2A (H2A) and H2AX, which in turn recruits DNA damage repair proteins, e.g., RAP80, BRCA1. In addition, SUMOylation and NEDDylation are also involved in the DNA damage response. RNF168 is a dual specificity E3 ligase that mediates both the ubiquitination and NEDDylation of H2A and H2AX. The two types of modifications compete against each other and NEDD8 modification of H2A and H2AX blocks the recruitment of BRCA1 at DNA damage repair sites. RNF168 itself is also subjected to NEDD8 modification and NEDDylation of RNF168 is necessary for its ubiquitin ligase activity. Inhibition of RNF168 NEDDylation impairs its interaction with E2 enzyme Ubc13 (UBE2N). Likewise, downregulating RNF168 NEDDylation by the deNEDDylating enzyme NEDP1, or due to mutations of the NEDD8-conjugating enzyme UBC12, decreases H2A and H2AX ubiquitylation. RNF168 Mutations are linked to RIDDLE (radiosensitivity, immunodeficiency dysmorphic features and learning difficulties) syndrome.

Immunogen

GST-tagged recombinant human RNF168.

Epitope: C-terminus

Other Notes

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

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.

Quality

Evaluated by Western Blotting in SA13 human B cell lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected RNF168 in 8 µg of SA13 human B cell lysate.

Specificity

This antibody detects RNF168 in a lymphoblastoid cell line (LCL) derived from a healthy donor, but not LCL derived from a RIDDLE (radiosensitivity, immunodeficiency dysmorphic features and learning difficulties) syndrome patient.

Storage and Stability

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

Target description

~75 kDa observed. Uncharacterized band(s) may appear in some lysates.