Product datasheet Anti-RNF20 antibody ab70495 1 Abreviews 1 References 3 Images Overview Product name Anti-RNF20 antibody Description Rabbit polyclonal to RNF20 Specificity We have data to indicate that this antibody may not cross react with Mouse. However, this has not been conclusively tested and expression levels may vary in certain cell lines/tissues. Tested applications WB, IP, ICC/IF Species reactivity Reacts with: Human Predicted to work with: Mouse, Rat, Rabbit, Dog, Pig, Chimpanzee, Ferret, Rhesus monkey, Gorilla, Chinese Hamster, Orangutan Immunogen Synthetic peptide, corresponding to a region between amino acids 1-50 of Human RNF20 Positive control Whole cell lysate from HeLa and 293T cells Properties Form Liquid Storage instructions Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. Storage buffer Preservative: 0.09% Sodium Azide Constituents: 8mM PBS, 60mM Citrate, 150mM Tris, pH 7 to 8 Purity Immunogen affinity purified Clonality Polyclonal Isotype IgG Applications Our Abpromise guarantee covers the use of ab70495 in the following tested applications. The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user. Application WB Abreviews Notes 1/2000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 114 kDa). IP Use at 2 µg/mg of lysate. ICC/IF 1/1000. 1 Target Function Component of the RNF20/40 E3 ubiquitin-protein ligase complex that mediates monoubiquitination of 'Lys-120' of histone H2B (H2BK120ub1). H2BK120ub1 gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation (H3K4me and H3K79me, respectively). It thereby plays a central role inb histone code and gene regulation. The RNF20/40 complex forms a H2B ubiquitin ligase complex in cooperation with the E2 enzyme UBE2A or UBE2B; reports about the cooperation with UBE2E1/UBCH are contradictory. Required for transcriptional activation of Hox genes. Recruited to the MDM2 promoter, probably by being recruited by p53/TP53, and thereby acts as a transcriptional coactivator. Mediates the polyubiquitination of isoform 2 of PA2G4 in cancer cells leading to its proteasome-mediated degradation. Tissue specificity Expressed in the normal brain and also in malignant gliomas (at protein level). Pathway Protein modification; protein ubiquitination. Sequence similarities Belongs to the BRE1 family. Contains 1 RING-type zinc finger. Cellular localization Nucleus. Anti-RNF20 antibody images All lanes : Anti-RNF20 antibody (ab70495) at 0.1 µg/ml Lane 1 : Whole cell lysate from HeLa cells at 50 µg Lane 2 : Whole cell lysate from HeLa cells at Western blot - RNF20 antibody (ab70495) 15 µg Lane 3 : Whole cell lysate from HeLa cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg developed using the ECL technique Predicted band size : 114 kDa Observed band size : 140 kDa Additional bands at : 171 kDa,280 kDa. We are unsure as to the identity of these extra bands. Exposure time : 30 seconds 2 Detection of Human RNF20 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded) using ab70495 at 3 µg/mg lysate. Subsequent RNF20 WB detection was performed using ab70495 at 1 µg/ml. Immunoprecipitation - RNF20 antibody (ab70495) ab70495 (1/1000) detecting RNF20 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to Abreview for further Immunocytochemistry/ Immunofluorescence - experimental details. RNF20 antibody (ab70495) This image is courtesy of an Abreview submitted by Dr Kirk McManus Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE" Our Abpromise to you: Quality guaranteed and expert technical support Replacement or refund for products not performing as stated on the datasheet Valid for 12 months from date of delivery Response to your inquiry within 24 hours We provide support in Chinese, English, French, German, Japanese and Spanish Extensive multi-media technical resources to help you We investigate all quality concerns to ensure our products perform to the highest standards If the product does not perform as described on this datasheet, we will offer a refund or replacement. For full details of the Abpromise, please visit http://www.abcam.com/abpromise or contact our technical team. Terms and conditions Guarantee only valid for products bought direct from Abcam or one of our authorized distributors 3