Product nameAnti-HDAC9 antibody [EPR5223]
See all HDAC9 primary antibodies
DescriptionRabbit monoclonal [EPR5223] to HDAC9
Tested applicationsSuitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
Unsuitable for: IP
Species reactivityReacts with: Rat, Human
Synthetic peptide within Human HDAC9 aa 1-100 (N terminal). The exact sequence is proprietary.
- IHC-P: Human colon tissue ICC/IF: HeLa cells WB: HepG2, Raji, HAP1, cell lysates FC: K-562 cells.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab109446 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent concentration.|
|WB||1/10000 - 1/50000. Detects a band of approximately 160 kDa (predicted molecular weight: 111 kDa).|
|IHC-P||1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||1/100 - 1/250.|
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.
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.
- Information by UniProt
- HD 7 antibody
- HD 7B antibody
- HD 9 antibody
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 specifically recognize HDAC9 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when HDAC9 knockout samples were usedexamined. Wild-type and HDAC9 knockout samples were subjected to SDS-PAGE. Ab109446 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10,000 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.
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
Lysates/proteins at 10 µg per lane.
Predicted band size: 111 kDa
Observed band size: 160 kDa why is the actual band size different from the predicted?
ab109446 at 1/250 dilution staining HDAC9 in Human colon by Immunohistochemistry, Paraffin-embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ab109446 at 1/100 dilution staining HDAC9 in HeLa cells by Immunofluorescence.
Flow cytometry of K-562 Human chronic myelogenous leukemia lymphoblast cells staining HDAC9 with ab109446 (red) at 1:250. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used at 1:2000 as a secondary. Rabbit monoclonal IgG (ab172730) (Black) was used as an isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) used as an unlabelled control. Fixed with 4% paraformaldehyde. Permeabilised with 90% methanol.
This product has been referenced in:
- Li A et al. Histone deacetylase 6 regulates the immunosuppressive properties of cancer-associated fibroblasts in breast cancer through the STAT3-COX2-dependent pathway. Oncogene N/A:N/A (2018). Read more (PubMed: 29980788) »
- Jin Q et al. MicroRNA-101-3p inhibits proliferation in retinoblastoma cells by targeting EZH2 and HDAC9. Exp Ther Med 16:1663-1670 (2018). Read more (PubMed: 30186385) »