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Immunofluorescence analysis of HDAC2 was done on 70% confluent log phase A431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HDAC2 Rabbit Polyclonal Antibody (515100) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human , Mouse|
|Published species reactivity||Mouse , Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic 11 amino acid peptide derived from the C-terminus of the mouse HDAC2 protein.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||Assay Dependent|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2-3 µg/mL|
|Immunofluorescence (IF)||2-3 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Immunoprecipitation (IP)||5 ug|
|Western Blot (WB)||1-3 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.
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.
Deacetylase inhibitors repress STAT5-mediated transcription by interfering with bromodomain and extra-terminal (BET) protein function.
51-5100 was used in ChIP assay and western blot to test if deacetylase inhibitors target STAT5 or histone acetylation
|Pinz S,Unser S,Buob D,Fischer P,Jobst B,Rascle A||Nucleic acids research (43:3524)||2015|
Redox-sensitive gene-regulatory events controlling aberrant matrix metalloproteinase-1 expression.
51-5100 was used in western blot to demonstrate that histone deacetylase-2 protein levels and its occupancy of the MMP-1 promoter are decreased in response to enforced manganese superoxide dismutase expression
|Bartling TR,Subbaram S,Clark RR,Chandrasekaran A,Kar S,Melendez JA||Free radical biology & medicine (74:99)||2014|
Transcriptional repression of ER through hMAPK dependent histone deacetylation by class I HDACs.
51-5100 was used in western blot to study the relationship between hyperactivation of MAPK and the estrogen receptor in breast cancer cells
|Plotkin A,Volmar CH,Wahlestedt C,Ayad N,El-Ashry D||Breast cancer research and treatment (147:249)||2014|
pVHL-mediated transcriptional repression of c-Myc by recruitment of histone deacetylases.
51-5100 was used in western blot to identify a novel mechanism for pVHL-mediated negative regulation of c-Myc transcription.
|Hwang IY,Roe JS,Seol JH,Kim HR,Cho EJ,Youn HD||Molecules and cells (33:195)||2012|
Heregulin regulates the ability of the ErbB3-binding protein Ebp1 to bind E2F promoter elements and repress E2F-mediated transcription.
||Zhang Y,Hamburger AW||The Journal of biological chemistry (279:26126)||2004|
Repression of E2F1-mediated transcription by the ErbB3 binding protein Ebp1 involves histone deacetylases.
||Zhang Y,Woodford N,Xia X,Hamburger AW||Nucleic acids research (31:2168)||2003|
Spatial re-organization of myogenic regulatory sequences temporally controls gene expression.
51-5100 was used in ChIP assay to show that the spatial organization of late genes contributes to temporal regulation of myogenic transcription during myogenesis
|Harada A,Mallappa C,Okada S,Butler JT,Baker SP,Lawrence JB,Ohkawa Y,Imbalzano AN||Nucleic acids research (43:2008)||2015|
Contrasting roles for MyoD in organizing myogenic promoter structures during embryonic skeletal muscle development.
51-5100 was used in ChIP assay, immunocytochemistry, immunoprecipitation, and western blot to study the temporal regulation of MyoD in skeletal muscle differentiation
|Cho OH,Mallappa C,Hernández-Hernández JM,Rivera-Pérez JA,Imbalzano AN||Developmental dynamics : an official publication of the American Association of Anatomists (244:43)||2015|
The histone deacetylase inhibitor valproic acid selectively induces proteasomal degradation of HDAC2.
51-5100 was used in immunohistochemistry - paraffin section to demonstrate that poly-ubiquitination and proteasomal degradation are an isoenzyme-selective mechanism for the downregulation of HDAC2.
|Krämer OH,Zhu P,Ostendorff HP,Golebiewski M,Tiefenbach J,Peters MA,Brill B,Groner B,Bach I,Heinzel T,Göttlicher M||The EMBO journal (22:3411)||2003|
mRPD3, HD2, D10Wsu179e, YAF1, Yy1bp, RPD3
YY1-associated factor 1, histone deacetylase 2, transcriptional regulator homolog RPD3, HD2, YY1 transcription factor-binding protein, HDAC2, Histone deacetylase 2, RPD3, YAF1