The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 71 kDa (predicted molecular weight: 71 kDa).
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Histone demethylase that specifically demethylates 'Lys-4' (H3K4me) and 'Lys-36' (H3K36me) of histone H3, thereby playing a central role in histone code. Preferentially demethylates trimethylated H3 'Lys-4' (H3K4me3) and monomethylated H3 'Lys-4' (H3K4me1) residues, while it has weaker activity for dimethylated H3 'Lys-36' (H3K36me2). Acts as a regulator of osteoblast differentiation via its interaction with SP7/OSX by demethylating H3K4me and H3K36me, thereby inhibiting SP7/OSX-mediated promoter activation (By similarity). May also play a role in ribosome biogenesis and in the replication or remodeling of certain heterochromatic region.
Belongs to the MINA53/NO66 family. NO66 subfamily. Contains 1 JmjC domain.
Nucleus > nucleolus. Nucleus > nucleoplasm. Granular part of nucleoli. Nucleoplasm, nucleoplasmic foci, some of them associated with nucleoli.
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human kidney tissue labeling C14orf169 / NO66 with ab194292 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelial cells of human colon. Counter-stained with Hematoxylin. Secondary antibody only control: PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunoprecipitation of C14orf169 / NO66 from 1mg of HEK293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab194292 at 1/1000 dilution. Western blot was performed from the immunoprecipitate using ab194292 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) at 1/10000 was used as secondary antibody. Lane 1: HEK293 whole cell lysate 10µg (Input). Lane 2: ab194292 IP in HEK293 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194292 in HEK293 whole cell lysates. Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 second.
Western blot - Anti-C14orf169 / NO66 antibody [EPR18754] (ab194292)
Lanes 1-4 : Anti-C14orf169 / NO66 antibody [EPR18754] (ab194292) at 1/1000 dilution Lane 5 : Anti-C14orf169 / NO66 antibody [EPR18754] (ab194292) at 1/5000 dilution
Lane 1 : HEK293 (Human epithelial cells from embryonic kidney) whole cell lysate Lane 2 : U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate Lane 3 : A549 (Human lung carcinoma) whole cell lysate Lane 4 : NCCIT (human pluripotent embryonic carcinoma) whole cell lysate Lane 5 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution