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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M | WB, ICC | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | ABE367 |
| Description | Anti-RNF168 Antibody |
| Alternate Names |
|
| Background Information | E3 ubiquitin-protein ligase RNF168 (UniProt: Q8IYW5; also known as EC:2.3.2.27, hRNF168, RING finger protein 168, RING-type E3 ubiquitin transferase RNF168) is encoded by the RNF168 gene (Gene ID: 165918) in human. RNF168 is a E3 ubiquitin-protein ligase that is required for accumulation of repair proteins, such as RAP80 and BRCA1 to sites of DNA damage. It acts with UBE2N/UBC13 to amplify the RNF8-dependent histone ubiquitination. It is recruited to sites of DNA damage at double-strand breaks (DSBs) by binding to ubiquitinated histone H2A and H2AX and amplifies the RNF8-dependent H2A ubiquitination, promoting the formation of lysine 63-linked ubiquitin conjugates. This leads to concentrate ubiquitinated histones H2A and H2AX at DNA lesions to the threshold required for recruitment of TP53BP1 and BRCA1. RNF168 can serve as a dual specificity E3 ligase to mediate 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 its NEDDylation is shown to be essential for its ubiquitin ligase activity. Inhibition of RNF168 NEDDylation is shown to impair 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 have been linked to RIDDLE syndrome that is characterized by increased radiosensitivity, immunodeficiency, mild motor control and learning difficulties, facial dysmorphism, and short stature. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
|
| Presentation | Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-RNF168, Cat. No. ABE367, is a rabbit polyclonal antibody that detects RNF168 and is tested for use in Immunocytochemistry and Western Blotting. |
| Key Applications |
|
| Application Notes | Tested Applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected RNF168 in HeLa cell lysate. Immunocytochemistry Analysis: A 1:200 dilution from a representative lot detected RNF168 in NIH3T3, A431, HUVEC, and HeLa cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Biological Information | |
|---|---|
| Immunogen | Full-length recombinant human E3 ubiquitin-protein ligase RNF168. |
| Epitope | C-terminus |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody detects E3 ubiquitin-protein ligase RNF168. |
| Species Reactivity |
|
| Species Reactivity Note | Human, Mouse. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
|
| Entrez Gene Summary | This gene encodes an E3 ubiquitin ligase protein that contains a RING finger, a motif present in a variety of functionally distinct proteins and known to be involved in protein-DNA and protein-protein interactions. The protein is involved in DNA double-strand break (DSB) repair. Mutations in this gene result in Riddle syndrome. [provided by RefSeq, Sep 2011]. |
| Gene Symbol |
|
| Purification Method | Affinity Purfied |
| UniProt Number |
|
| UniProt Summary | FUNCTION: E3 ubiquitin-protein ligase required for accumulation of repair proteins to sites of DNA damage. Acts with UBE2N/UBC13 to amplify the RNF8-dependent histone ubiquitination. Recruited to sites of DNA damage at double-strand breaks (DSBs) by binding to ubiquitinated histone H2A and ubiquitinates histone H2A and H2AX, leading to amplify the RNF8-dependent H2A ubiquitination and promoting the formation of Lys-63-linked ubiquitin conjugates. This leads to concentrate ubiquitinated histones H2A and H2AX at DNA lesions to the threshold required for recruitment of TP53BP1 and BRCA1. PATHWAY: Protein modification; protein ubiquitination. SUBUNIT STRUCTURE: Interacts with UBE2N/UBC13. SUBCELLULAR LOCATION: Nucleus. Note: Localizes to sites of DNA damage. DOMAIN: The MIU motifs (motif interacting with ubiquitin) mediate the interaction with ubiquitin and the localization at sites of DNA damage. INVOLVEMENT IN DISEASE: Defects in RNF168 are the cause of Riddle syndrome (RIDDLES) [MIM:611943]. Riddle syndrome is characterized by increased radiosensitivity, immunodeficiency, mild motor control and learning difficulties, facial dysmorphism, and short stature. Defects are probably due to impaired localization of TP53BP1 and BRCA1 at DNA lesions. SEQUENCE SIMILARITIES: Belongs to the RNF168 family. Contains 1 RING-type zinc finger. SEQUENCE CAUTION: The sequence BAB70801.1 differs from that shown. Reason: Erroneous initiation. The sequence BAC04060.1 differs from that shown. Reason: Erroneous initiation. |
| Molecular Weight | ~75 kDa observed, 65.02 kDa calculated. Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in SA13 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected RNF168 in SA13 cell lysate. |
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Recommend storage at +2°C to +8°C. For long term storage antibodies can be kept at -20°C. Avoid repeated freeze-thaws. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |