Human RNF8 Antibody

Catalog #: AF7114 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF7114
AF7114-SP

Detection of Human RNF8 by Western Blot.

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Human RNF8 Antibody Summary

Species Reactivity

Human

Specificity

Detects human RNF8 in direct ELISAs and Western blots. In direct ELISAs, approximately 100% cross-reactivity with recombinant mouse RNF8 is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human RNF8
Met1-Val124
Accession # O76064

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.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

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

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Detection of Human RNF8 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human RNF8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7114) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for RNF8 at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

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: RNF8

RNF8 (RING [really interesting gene] finger protein 8; also E3 ubiquitin-protein ligase RNF8) is a 55-58 kDa member of the RNF8 family of proteins. It is ubiquitously expressed, and serves as an E3 ubiquitin ligase. RNF8 is recruited to DNA double-strand breaks by phosphoMDC1, where it promotes the ubiquitination of histones H2A and H2AX. This, in turn, may result in the recruitment of 53BP1, a scaffold protein that holds DNA damage response elements. Human RNF8 is 485 amino acids (aa) in length. It contains one forkhead associated domain (aa 38-92), a Gln-rich segment (aa 276-345), and one Zn-finger region (aa 403-441). There is one utilized phosphorylation site at Ser157. Two potential isoforms are reported. One shows an 18 aa substitution for aa 81-485, while another possesses a 36 aa substitution for aa 413-485. RNF8 apparently undergoes ubiquitination, generating multiple isoforms that run at 62-72 kDa in SDS-Page. Over aa 1-124, human RNF8 shares 74% aa identity with mouse RNF8.

Long Name

Ring Finger Protein 8

Entrez Gene IDs

9025 (Human); 58230 (Mouse)

Alternate Names

ActA Binding Protein 2; C3HC4-type zinc finger protein; EC 6.3.2; EC 6.3.2.-; FLJ12013; KIAA0646UBC13/UEV-interacting ring finger protein; ring finger protein (C3HC4 type) 8; ring finger protein 8E3 ubiquitin-protein ligase RNF8; RNF8