The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10000 - 1/50000. Detects a band of approximately 160 kDa (predicted molecular weight: 111 kDa).
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/100 - 1/250.
Application notesIs unsuitable for Flow Cyt or IP.
FunctionResponsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Represses MEF2-dependent transcription. Isoform 3 lacks active site residues and therefore is catalytically inactive. Represses MEF2-dependent transcription by recruiting HDAC1 and/or HDAC3. Seems to inhibit skeletal myogenesis and to be involved in heart development. Protects neurons from apoptosis, both by inhibiting JUN phosphorylation by MAPK10 and by repressing JUN transcription via HDAC1 recruitment to JUN promoter.
Tissue specificityBroadly expressed, with highest levels in brain, heart, muscle and testis. Isoform 3 is present in human bladder carcinoma cells (at protein level).
Involvement in diseaseNote=A chromosomal aberration involving HDAC9 is found in a family with Peters anomaly. Translocation t(1;7)(q41;p21) with TGFB2 resulting in lack of HDAC9 protein.
Sequence similaritiesBelongs to the histone deacetylase family. HD type 2 subfamily.
Post-translational modificationsPhosphorylated on Ser-220 and Ser-450; which promotes 14-3-3-binding, impairs interaction with MEF2, and antagonizes antimyogenic activity. Phosphorylated on Ser-240; which impairs nuclear accumulation (By similarity). Isoform 7 is phosphorylated on Tyr-1010. Phosphorylated by the PKC kinases PKN1 and PKN2, impairing nuclear import. Sumoylated.
Lane 1: Wild type HAP1 whole cell lysate (20 µg) Lane 2: HDAC9 (KO) knockout HAP1 whole cell lysate (20 µg) Lane 3: HepG2 whole cell lysate (20 µg) Lane 4: Raji whole cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab109446 observed at 140 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109446 was shown to recognize HDAC9 (KO) when HDAC9 (KO) knockout samples were used, along with additional cross-reactive bands. Wild-type and HDAC9 (KO) knockout samples were subjected to SDS-PAGE. Ab109446 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 10000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - HDAC9 antibody [EPR5223] (ab109446)
All lanes : Anti-HDAC9 antibody [EPR5223] (ab109446) at 1/10000 dilution
Lane 1 : K562 cell lysate Lane 2 : HepG2 cell lysate Lane 3 : Raji cell lysate
ab109446 at 1/100 dilution staining HDAC9 in HeLa cells by Immunofluorescence.
References for Anti-HDAC9 antibody [EPR5223] (ab109446)
This product has been referenced in:
Di Giorgio E et al. The co-existence of transcriptional activator and transcriptional repressor MEF2 complexes influences tumor aggressiveness. PLoS Genet13:e1006752 (2017).
Read more (PubMed: 28419090) »
Sebastian S et al. Tissue-specific splicing of a ubiquitously expressed transcription factor is essential for muscle differentiation. Genes Dev27:1247-59 (2013).
Read more (PubMed: 23723416) »