1/200. Use with methanol or paraformaldehyde fixed cells.
Use at an assay dependent concentration.
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
FunctionE3 ubiquitin-protein ligase that mediates monoubiquitination of 'Lys-119' of histone H2A, thereby playing a central role in histone code and gene regulation. H2A 'Lys-119' ubiquitination gives a specific tag for epigenetic transcriptional repression and participates in X chromosome inactivation of female mammals. May be involved in the initiation of both imprinted and random X inactivation. Essential component of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex act via chromatin remodeling and modification of histones, rendering chromatin heritably changed in its expressibility. E3 ubiquitin-protein ligase activity is enhanced by BMI1/PCGF4. Acts as the main E3 ubiquitin ligase on histone H2A of the PRC1 complex, while RING1 may rather act as a modulator of RNF2/RING2 activity.
PathwayProtein modification; protein ubiquitination.
Post-translational modificationsPolyubiquitinated in the presence of UBE2D3 (in vitro). Monoubiquitinated, by auto-ubiquitination.
Cellular localizationNucleus. Chromosome. Enriched on inactive X chromosome (Xi) in female trophoblast stem (TS) cells as well as differentiating embryonic stem (ES) cells. The enrichment on Xi is transient during TS and ES cell differentiation. The association with Xi is mitotically stable in non-differentiated TS cells.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling RING2 / RING1B / RNF2 with ab101273 at 1/1000 (1µg/ml).
Western blot - RING2 / RING1B / RNF2 antibody (ab101273)
All lanes : Anti-RING2 / RING1B / RNF2 antibody - ChIP Grade (ab101273) at 0.1 µg/ml
Lane 1 : K562 whole cell lysate at 50 µg Lane 2 : K562 whole cell lysate at 15 µg Lane 3 : K562 whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg Lane 5 : NIH3T3 whole cell lysate at 50 µg
Detection of RING2 / RING1B / RNF2 in Immunoprecipitates of K562 whole cell lysate (1 mg for IP, 20% of IP loaded) using ab101273 at 3 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection. Detection: Chemiluminescence with an exposure time of 30 seconds.
Immunocytochemistry/ Immunofluorescence - RING2 / RING1B / RNF2 antibody (ab101273)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
ab101273 (1/200) staining RING2 in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.