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Human/Mouse RNF168 Antibody

Catalog #: AF7217
6 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF7217
AF7217-SP
Detection of Mouse RNF168 by Western Blot.
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Citations (6)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse RNF168 Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects recombinant mouse RNF168 and recombinant human RNF168 in direct ELISAs and Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant mouse RNF168
Asn423-Arg565
Accession # Q80XJ2
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
Immunocytochemistry
This antibody has been used at a concentration of 5-15 μg/mL to detect RNF168 in immersion fixed HeLa human cervical epithelial carcinoma cell line. Internal testing was not able to validate staining in a mouse cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Mouse RNF168 antibody by Western Blot. View Larger

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.

Immunocytochemistry RNF168 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

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; NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunocytochemistry View Larger

Detection of RNF168 in RAW 264.7 Mouse Monocyte/Macrophage Cell Line. RNF168 was detected in immersion fixed RAW 264.7 Mouse Monocyte/Macrophage Cell Line using Sheep Anti-Human/Mouse RNF168 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7217) at 5 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). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: RNF168

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.

Long Name
Ring Finger Protein 168
Entrez Gene IDs
165918 (Human); 70238 (Mouse)
Alternate Names
E3 ubiquitin-protein ligase RNF168; EC 6.3.2; EC 6.3.2.-; FLJ39749; ring finger protein 168FLJ35794; RNF168

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Citations for Human/Mouse RNF168 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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  1. RNF168-mediated localization of BARD1 recruits the BRCA1-PALB2 complex to DNA damage
    Authors: John J. Krais, Yifan Wang, Pooja Patel, Jayati Basu, Andrea J. Bernhardy, Neil Johnson
    Nature Communications
  2. Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling
    Authors: Longarini EJ, Dauben H, Locatelli C et al.
    Molecular cell
  3. RNF168-mediated ubiquitin signaling inhibits the viability of BRCA1 null cancers
    Authors: JJ Krais, Y Wang, AJ Bernhardy, E Clausen, JA Miller, KQ Cai, CL Scott, N Johnson
    Cancer Res., 2020-03-25;0(0):.
  4. RNF168 and USP10 regulate topoisomerase II? function via opposing effects on its ubiquitylation
    Nat Commun, 2016-08-25;7(0):12638.
    Species: Human
    Sample Types: Whole Cells
    Applications: IHC-Fr
  5. RNF168 regulates R-loop resolution and genomic stability in BRCA1/2-deficient tumors
    Authors: Parasvi S. Patel, Karan Joshua Abraham, Kiran Kumar Naidu Guturi, Marie-Jo Halaby, Zahra Khan, Luis Palomero et al.
    Journal of Clinical Investigation
  6. 53BP1 Enforces Distinct Pre- and Post-resection Blocks on Homologous Recombination
    Authors: Callen E, Zong D, Wu W et al.
    Mol. Cell

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