Invitrogen

HDAC2 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

8 Published Figures

14 References

View all (41) HDAC2 antibodies

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Datasheet

Protocols

Questions & Answers

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HDAC2 Antibody in Immunocytochemistry (ICC/IF) — 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 1 µg/mL for 30 minutes. Images were taken on a Zeiss Axio Observer Z1 microscope at 20X magnification.

HDAC2 Antibody in ChIP Assay (ChIP) — Chromatin immunoprecipitation analysis of HDAC2 was performed using cross-linked chromatin from 1x10^6 HCT116 colon carcinoma cells treated with serum for 0, 15, and 30 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of an HDAC2 polyclonal antibody (Product # PA1-861). Chromatin aliquots from ~1x10^5 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify -15kb upstream of the Egr1 gene or exon-1 or exon-2 of Egr1. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the Egr-1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars. Data courtesy of the Innovators Program.

HDAC2 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on cancer biopsies of deparaffinized human esophagus carcinoma. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a Rabbit Polyclonal Antibody recognizing HDAC2 (Product # PA1-861) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

HDAC2 Antibody in Western Blot (WB) — Western blot analysis of HDAC2 was performed by loading HeLa cell lysate onto a SDS-PAGE gel. Proteins were transferred to a membrane, blocked, and probed with a HDAC2 polyclonal antibody (Product # PA1-861) at a concentration of 0.5 µg/mL followed by an HRP-conjugated anti-rabbit IgG secondary antibody. Detection was performed using a chemiluminescent substrate.

HDAC2 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of HDAC2 was done on HeLa cells. Cells were fixed, permeabilized and stained with a HDAC2 rabbit polyclonal antibody (Product # PA1-861, red histogram) or a rabbit IgG isotype control (Product # MA5-16384, black histogram) at a dilution of 10 µg/mL. After incubation for 1 hour on ice, the cells were labeled with a Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 647 conjugate (Product # A27040) at a dilution of 1:50 for 1 hour on ice. A representative 10,000 cells were acquired and analyzed for each sample.

HDAC2 Antibody in Immunoprecipitation (IP) — Figure 9. Identify the binding of the HAT & HDAC to the TH promoter by ChIP assay (all N = 4/group). Exposure of MN9D cells to NE for two days increased CBP/p300 binding to TH promoter (A) and reduced the binding of HDAC2 (B) and HDAC5 (C) to the TH promoter. ""Input"" serves as a loading control and rabbit IgG immunoprecipitation serves as a negative control. Top panel in each figures showed the binding to the TH promoter in response to NE treatment. Low panel in each figure showed quantitative real-time PCR of the TH promoter regions from immunoprecipitation with antibody against CBP/p300, HDAC2, and HDAC5. The fold enrichment value is shown as the normalized ChIP signals divided by the normalized input signal. Each bar from both pictures represent data obtained from four separate experiments ( N = 4/group). ** p < .01, compared to the control. NE: treatment with 300 nM NE. Abbreviations: ChIP: chromatin immunoprecipitation assay; HAT: histone acetyl transferase; HDAC: histone deacetylases; NE: norepinephrine; PCR: polymerase chain reaction; TH: tyrosine hydroxylase.

HDAC2 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein inHDAC2 KO cell line compared to control cell line using Anti-HDAC2 Polyclonal Antibody (Product # PA1-861). {KO}

Product Details

PA1-861

Applications	Tested Dilution	Publications
Western Blot (WB)	0.5 µg/mL	View 11 publications 11 publications
Immunohistochemistry (IHC)	0.4 µg/mL	-
Immunocytochemistry (ICC/IF)	1.0 µg/mL	View 1 publication 1 publication
Flow Cytometry (Flow)	1-20 µg/mL	-
Immunoprecipitation (IP)	Assay-dependent	View 1 publication 1 publication
ChIP assay (ChIP)	1-3 µL	View 1 publication 1 publication

Product Specifications
Species Reactivity	Dog, Hamster, Human, Mouse, Non-human primate, Rat
Published species	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
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. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2118520

Product Specific Information

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.

Target Information

HDAC2 belongs to the histone deacetylase family. Histone deacetylases act via the formation of large multiprotein complexes and are responsible for the deacetylation of lysine residues at the N-terminal regions of core histones (H2A, H2B, H3 and H4). This protein forms transcriptional repressor complexes by associating with many different proteins, including YY1, a mammalian zinc-finger transcription factor. Thus, it plays an important role in transcriptional regulation, cell cycle progression and developmental events. Alternative splicing results in multiple transcript variants.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Effects of Manipulation of Noradrenergic Activities on the Expression of Dopaminergic Phenotypes in Aged Rat Brains.

ASN neuro

Zeng F,Fan Y,Brown RW,Drew Gill W,Price JB,Jones TC,Zhu MY

Published figure using HDAC2 polyclonal antibody (Product # PA1-861) in Immunoprecipitation

Mon Jan 17 00:00:00 EST 2022

Neuronal activity-induced BRG1 phosphorylation regulates enhancer activation.

Cell reports

Kim B,Luo Y,Zhan X,Zhang Z,Shi X,Yi J,Xuan Z,Wu J

PA1-861 was used in Western Blot to study how BRG1 regulates enhancer basal activities and inducibility by affecting cohesin binding, enhancer-promoter looping, RNA polymerase II recruitment, and enhancer RNA expression. Using mice with knockin mutations, to show that non-phosphorylatable BRG1 fails to efficiently induce activity-dependent genes, whereas phosphomimic BRG1 increases enhancer activity and inducibility.

Tue Jul 13 00:00:00 EDT 2021

Loss of Asxl2 leads to myeloid malignancies in mice.

Nature communications

Li J,He F,Zhang P,Chen S,Shi H,Sun Y,Guo Y,Yang H,Man N,Greenblatt S,Li Z,Guo Z,Zhou Y,Wang L,Morey L,Williams S,Chen X,Wang QT,Nimer SD,Yu P,Wang QF,Xu M,Yang FC

PA1-861 was used in Western Blotting to demonstrate that ASXenopus laevis2 functions as a tumour suppressor to maintain normal haematopoietic stem cell function.

Thu Jun 08 00:00:00 EDT 2017

Class I lysine deacetylases promote glucocorticoid-induced transcriptional repression through functional interaction with LSD1.

The Journal of steroid biochemistry and molecular biology

Patrick NM,Griggs CA,Icenogle AL,Gilpatrick MM,Kadiyala V,Jaime-Frias R,Smith CL

PA1-861 was used in Western Blotting to demonstrate that Class I lysine deacetylases facilitate dexamethasone-induced transcriptional repression by suppressing LSD1 complex activity at selected target gene promoters.

Wed Mar 01 00:00:00 EST 2017

Histone deacetylases meet microRNA-associated MMP-9 expression regulation in glucocorticoid-sensitive and -resistant cell lines.

International journal of oncology

Hentati-Kallel M,Le Jan S,Bernard P,Antonicelli F,Trussardi-Régnier A

PA1861 was used in western blot to suggest that the use of HDAC inhibitors in a context of acquired glucocorticoid resistance may modify the epigenetic landscape

Wed Feb 01 00:00:00 EST 2017

Nuclear LC3 Associates with Slowly Diffusing Complexes that Survey the Nucleolus.

Traffic (Copenhagen, Denmark)

Kraft LJ,Manral P,Dowler J,Kenworthy AK

PA1-861 was used in western blot to test nuclear LC3 and how it associates with slowly diffusing complexes that interact with the nucleolus

Fri Apr 01 00:00:00 EDT 2016

RNA-dependent dynamic histone acetylation regulates MCL1 alternative splicing.

Nucleic acids research

Khan DH,Gonzalez C,Cooper C,Sun JM,Chen HY,Healy S,Xu W,Smith KT,Workman JL,Leygue E,Davie JR

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

Sat Feb 01 00:00:00 EST 2014

Protein kinase CK2 regulates the dimerization of histone deacetylase 1 (HDAC1) and HDAC2 during mitosis.

The Journal of biological chemistry

Khan DH,He S,Yu J,Winter S,Cao W,Seiser C,Davie JR

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

Fri Jun 07 00:00:00 EDT 2013

Proteomic and functional analyses reveal the role of chromatin reader SFMBT1 in regulating epigenetic silencing and the myogenic gene program.

The Journal of biological chemistry

Lin S,Shen H,Li JL,Tang S,Gu Y,Chen Z,Hu C,Rice JC,Lu J,Wu L

PA1-861 was used in western blot to study the mechanism by which SFMBT1 modulates epigenetic silencing and mitogenic gene expression

Fri Mar 01 00:00:00 EST 2013

Transcriptional repression of peroxisome proliferator-activated receptor beta/delta in murine keratinocytes by CCAAT/enhancer-binding proteins.

The Journal of biological chemistry

Di-Poï N,Desvergne B,Michalik L,Wahli W

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.

Fri Nov 18 00:00:00 EST 2005

Transcriptional repression of peroxisome proliferator-activated receptor beta/delta in murine keratinocytes by CCAAT/enhancer-binding proteins.

The Journal of biological chemistry

Di-Poï N,Desvergne B,Michalik L,Wahli W

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.

Fri Nov 18 00:00:00 EST 2005

Differential intranuclear organization of transcription factors Sp1 and Sp3.

Molecular biology of the cell

He S,Sun JM,Li L,Davie JR

PA1-861 was used in immunocytochemistry to study the nuclear localization of transcription factors Sp1 and Sp3.

Thu Sep 01 00:00:00 EDT 2005

Differential intranuclear organization of transcription factors Sp1 and Sp3.

Molecular biology of the cell

He S,Sun JM,Li L,Davie JR

PA1-861 was used in immunocytochemistry to study the nuclear localization of transcription factors Sp1 and Sp3.

Thu Sep 01 00:00:00 EDT 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.

Nucleic acids research

Ling Y,Sankpal UT,Robertson AK,McNally JG,Karpova T,Robertson KD

PA1-861 was used in western blot to study the regulation of DNA methyltransferase Dnmt3a enzymatic activity and transcriptional regulation.

Wed Apr 21 00:00:00 EDT 2004

Ligand-induced recruitment of a histone deacetylase in the negative-feedback regulation of the thyrotropin beta gene.

The EMBO journal

Sasaki S,Lesoon-Wood LA,Dey A,Kuwata T,Weintraub BD,Humphrey G,Yang WM,Seto E,Yen PM,Howard BH,Ozato K

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

Fri Oct 01 00:00:00 EDT 1999

Bioinformatics

Protein Aliases: HD2; hdac 2; hdac-2; Histone deacetylase 2; histone deacetylase 2-like protein; Protein deacylase HDAC2; transcriptional regulator homolog RPD3; YY1 transcription factor-binding protein; YY1-associated factor 1

Gene Aliases: D10Wsu179e; HD2; HDAC2; I79_010219; mRPD3; RPD3; YAF1; Yy1bp

UniProt ID: (Human) Q92769, (Mouse) P70288

Entrez Gene ID: (Human) 3066, (Dog) 475035, (Mouse) 15182, (Rat) 84577, (Chinese hamster) 100764557

Function(s)

Process(es)

RNA polymerase II core promoter proximal region sequence-specific DNA binding RNA polymerase II distal enhancer sequence-specific DNA binding core promoter binding RNA polymerase II repressing transcription factor binding chromatin binding histone deacetylase activity protein binding transcription factor binding deacetylase activity enzyme binding heat shock protein binding nucleosomal DNA binding NAD-dependent histone deacetylase activity (H3-K14 specific) protein deacetylase activity sequence-specific DNA binding poly(A) RNA binding NF-kappaB binding transcription factor activity, sequence-specific DNA binding hydrolase activity histone deacetylase activity (H3-K14 specific) chromatin DNA binding histone deacetylase activity (H3-K9 specific) histone deacetylase activity (H4-K16 specific) Krueppel-associated box domain binding NAD-dependent histone deacetylase activity (H3-K9 specific) NAD-dependent histone deacetylase activity (H4-K16 specific) NAD-dependent histone deacetylase activity (H3-K18 specific)

HDAC2 Polyclonal Antibody

Product Details

PA1-861

Western Blot (WB)

Immunohistochemistry (IHC)

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Immunoprecipitation (IP)

ChIP assay (ChIP)

Species Reactivity

Published species

Host/Isotype

Class

Type

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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