Product nameAnti-HDAC2 antibody [3F3]
See all HDAC2 primary antibodies
DescriptionMouse monoclonal [3F3] to HDAC2
Specificityab51832 recognizes HDAC2.
Tested applicationsSuitable for: Flow Cyt, IHC-P, WB, IP, ChIP, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow
Synthetic peptide, corresponding to amino acids 473-488 of Human HDAC2
- Nuclear extract of HeLa cells and HAP1 whole cell lysate . IF/ICC: MCF7 cell line.
This antibody clone is manufactured by Abcam.
This monoclonal antibody to HDAC2 has been knockout validated in Western blot. The expected band for HDAC2 was observed in wild type cells and the band was not seen in HDAC2 knockout cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Concentration information loading...
PurityProtein G purified
ChIP Related Products
Our Abpromise guarantee covers the use of ab51832 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 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration.|
|WB||1/1000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).|
|IP||Use at an assay dependent concentration.
Use 10ug for 10-50ug cell lysate.
|ChIP||Use at an assay dependent concentration.
Use 10ug for 25ug of DNA
|ICC/IF||1/25 - 1/100.|
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. Histone deacetylases act via the formation of large multiprotein complexes.
Forms transcriptional repressor complexes by associating with MAD, SIN3, YY1 and N-COR. Interacts in the late S-phase of DNA-replication with DNMT1 in the other transcriptional repressor complex composed of DNMT1, DMAP1, PCNA, CAF1. Deacetylates TSHZ3 and regulates its transcriptional repressor activity.
Tissue specificityWidely expressed; lower levels in brain and lung.
Sequence similaritiesBelongs to the histone deacetylase family. HD type 1 subfamily.
modificationsS-nitrosylated by GAPDH. In neurons, S-Nitrosylation at Cys-262 and Cys-274 does not affect the enzyme activity but abolishes chromatin-binding, leading to increases acetylation of histones and activate genes that are associated with neuronal development. In embryonic cortical neurons, S-Nitrosylation regulates dendritic growth and branching.
- Information by UniProt
- D10Wsu179e antibody
- HD 2 antibody
- HD2 antibody
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: HDAC2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: A431 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab51832 observed at 65 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab51832 detected the expected band for HDAC2 in wild-type HAP1 cells and the band was not seen in HDAC2 knockout HAP1 cells. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab51832 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
ab51832 stained MCF7 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51832 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Overlay histogram showing HeLa cells stained with ab51832 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab51832, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
All lanes : Anti-HDAC2 antibody [3F3] (ab51832) at 5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 5 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lane 6 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?
Exposure time: 16 minutes
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab51832 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Chromatin was prepared from the cell lysate of rat liver, cultured hepatoma cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The primary antibody was diluted 1/100 and incubated with the sample for 1.5 hours at 25°C. Anti RNA polymerase II was used on the positive control, whilst normal mouse IgG was used in the negative control, to compare difference of HDAC2 binding on silencer DNA and promoter DNA of a gene. Data is normalized to 10% Input. The immunoprecipitated DNA was quantified by real time PCR.
This product has been referenced in:
- Hou X et al. Suppression of HDAC2 in Spinal Cord Alleviates Mechanical Hyperalgesia and Restores KCC2 Expression in a Rat Model of Bone Cancer Pain. Neuroscience 377:138-149 (2018). WB, IHC-P ; Rat . Read more (PubMed: 29482000) »
- Lin YH et al. Opening a New Time Window for Treatment of Stroke by Targeting HDAC2. J Neurosci 37:6712-6728 (2017). Read more (PubMed: 28592694) »