Immunofluorescent analysis of Phalloidin (magenta) and HDAC2 (green) in A431 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA (Product # 37525) in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a HDAC2 polyclonal antibody (Product # PA1-861) at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 633 Phalloidin (Product # 21840) at a dilution of 1:120 (2.5units/ml final concentration) and nuclei (blue) were stained with Hoechst (Product # 62249) at a concentration of 1ug/ml for 30 minutes. Images were taken on a Zeiss Axio Observer Z1 microscope at 20X magnification.
|Tested species reactivity||Dog, Hamster, Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C T(475) D T K G T K S E Q L S N P(488) of human HDAC2.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|Flow Cytometry (Flow)||1-20 µg/ml|
|Immunocytochemistry (ICC)||1.0 µg/ml|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunohistochemistry (IHC)||0.4 µg/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||0.5 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-861 detects histone deacetylase 2 (HDAC2) from canine, hamster, human, mouse, non-human primate, rat tissues and cells.
PA1-861 has been successfully used in Western blot, ChIP, immunoprecipitation, immunofluorescence, flow cytometry, immunocytochemistry and immunohistochemistry procedures. By Western blot, this antibody detects an ~55 kDa protein representing HDAC2 and a ~40 kDa protein of unknown identity from HeLa cell lysate. Immunocytochemical staining of HDAC2 in HeLa cells with PA1-861 results in nuclear staining.
The PA1-861 immunogen is a synthetic peptide corresponding to residues C T(475) D T K G T K S E Q L S N P(488) of human HDAC2. This immunizing peptide (Cat. # PEP-070) is available for use in neutralization and control experiments.
Histone deacetylase 2 (HDAC2), or transcriptional regulator homolog RPD3 L1, is highly homologous to the yeast transcription factor RPD3 (reduced potassium dependency 3) gene. As in yeast, human HDA2 is likely to be involved in regulating chromatin structure during transcription. It has been implicated to associate with YY1, a mammalian zinc-finger transcription factor, which negatively regulates transcription by tethering RPD3 to DNA as a cofactor. This process is highly concerved from yeast to human.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Nuclear LC3 Associates with Slowly Diffusing Complexes that Survey the Nucleolus.
PA1-861 was used in western blot to test nuclear LC3 and how it associates with slowly diffusing complexes that interact with the nucleolus
|Kraft LJ,Manral P,Dowler J,Kenworthy AK||Traffic (Copenhagen, Denmark) (17:369)||2016|
Protein kinase CK2 regulates the dimerization of histone deacetylase 1 (HDAC1) and HDAC2 during mitosis.
PA1-861 was used in western blot to study the modulation of the dimerization of HDAC-1 and -2 by CK2-mediated phosphorylation during mitosis
|Khan DH,He S,Yu J,Winter S,Cao W,Seiser C,Davie JR||The Journal of biological chemistry (288:16518)||2013|
Proteomic and functional analyses reveal the role of chromatin reader SFMBT1 in regulating epigenetic silencing and the myogenic gene program.
PA1-861 was used in western blot to study the mechanism by which SFMBT1 modulates epigenetic silencing and mitogenic gene expression
|Lin S,Shen H,Li JL,Tang S,Gu Y,Chen Z,Hu C,Rice JC,Lu J,Wu L||The Journal of biological chemistry (288:6238)||2013|
Transcriptional repression of peroxisome proliferator-activated receptor beta/delta in murine keratinocytes by CCAAT/enhancer-binding proteins.
PA1-861 was used in western blot to study the relationship between PPARbeta and C/EBP transcription factors and its effect on cell proliferation and differentiation.
|Di-Poï N,Desvergne B,Michalik L,Wahli W||The Journal of biological chemistry (280:38700)||2005|
Modification of de novo DNA methyltransferase 3a (Dnmt3a) by SUMO-1 modulates its interaction with histone deacetylases (HDACs) and its capacity to repress transcription.
PA1-861 was used in western blot to study the regulation of DNA methyltransferase Dnmt3a enzymatic activity and transcriptional regulation.
|Ling Y,Sankpal UT,Robertson AK,McNally JG,Karpova T,Robertson KD||Nucleic acids research (32:598)||2004|
Ligand-induced recruitment of a histone deacetylase in the negative-feedback regulation of the thyrotropin beta gene.
PA1-861 was used in western blot to investigate the influence of histone deacetylases on the negative-feedback regulation of thyroid-stimulating hormone beta gen
|Sasaki S,Lesoon-Wood LA,Dey A,Kuwata T,Weintraub BD,Humphrey G,Yang WM,Seto E,Yen PM,Howard BH,Ozato K||The EMBO journal (18:5389)||1999|
RNA-dependent dynamic histone acetylation regulates MCL1 alternative splicing.
PA1-861 was used in ChIP assay, immunoprecipitation, and western blot to study the modulation of MCL1 alternative splicing by an RNA-dependent mechanism involving dynamic histone acetylation
|Khan DH,Gonzalez C,Cooper C,Sun JM,Chen HY,Healy S,Xu W,Smith KT,Workman JL,Leygue E,Davie JR||Nucleic acids research (42:1656)||2014|
Differential intranuclear organization of transcription factors Sp1 and Sp3.
PA1-861 was used in immunocytochemistry to study the nuclear localization of transcription factors Sp1 and Sp3.
|He S,Sun JM,Li L,Davie JR||Molecular biology of the cell (16:4073)||2005|