The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml.
Use a concentration of 0.5 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 55.3 kDa).
Use at an assay dependent concentration.
機能Responsible 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.
組織特異性Widely expressed; lower levels in brain and lung.
配列類似性Belongs to the histone deacetylase family. HD type 1 subfamily.
翻訳後修飾S-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.
ICC/IF image of ab16032 stained HeLa cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab16032 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h 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.
Western blot - Anti-HDAC2 antibody (ab16032)
All lanes : Anti-HDAC2 antibody (ab16032) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution Developed using the ECL technique
HDAC2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5ug of Rabbit polyclonal to HDAC2 and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extractdiluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab16032. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 60ka: HDAC2.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HDAC2 antibody (ab16032)This image is courtesy of an abreview submitted by Antibody Solutions Ltd.
ab16032 (1/250) staining HDAC2 in paraffin-embedded Human tonsil tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit/mouse conjugated to HRP. For further experimental details please refer to abreview.
Exposure time : 30 secondsab16032 cross-reacts with Mouse 3T3 cells but shows no cross-reactivity against either Rat liver or Chicken cell lysates.
Anti-HDAC2 antibody (ab16032) 使用論文
This product has been referenced in:
Yao ZG et al. Regional and cell-type specific distribution of HDAC2 in the adult mouse brain. Brain Struct Funct218:563-73 (2013).
Read more (PubMed: 22532304) »
Lee TK et al. An N-terminal truncated carboxypeptidase E splice isoform induces tumor growth and is a biomarker for predicting future metastasis in human cancers. J Clin Invest : (2011).
Read more (PubMed: 21285511) »