Invitrogen

HDAC1 Polyclonal Antibody

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

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HDAC1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of HDAC1 was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with HDAC1 Polyclonal Antibody (Product # PA1-41120) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant 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 (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

HDAC1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of HDAC1 in Human testis. Samples were incubated in HDAC1 polyclonal antibody (Product # PA1-41120) using a dilution of 1:250. Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Staining was performed by Histowiz.

HDAC1 Antibody in Western Blot (WB) — Knockout of HDAC1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1087187_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of HDAC1 was performed by loading 30 µg of HeLa Wild type (Lane 1), HeLa Cas9 (Lane 2) and HeLa HDAC1 KO (Lane 3) modified whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-HDAC1 Polyclonal Antibody (Product # PA1-41120, 2 µg/mL dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to HDAC1. Uncharacterized band was observed in all the samples at 51 kDa.

HDAC1 Antibody in Western Blot (WB) — Western blot was performed using Anti-HDAC1 Polyclonal Antibody (Product # PA1-41120) and ~55kDa band corresponding to HDAC1 was observed in MCF-7, HeLa, HEK-293 and NIH/3T3 cells. Modified whole cell extracts (1% SDS) (30 ug lysate) of MCF-7 (Lane 1), HeLa (Lane 2), HEK-293 (Lane 3) and NIH/3T3 (Lane 4) were electrophoresed using NuPAGE® 4-12% Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001).The blot was probed with the primary antibody (2µg/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..

HDAC1 Antibody in Western Blot (WB) — Knockdown of HDAC1 was achieved by transfecting MCF7 cells with HDAC1 specific siRNAs (Silencer® select Product # s73). Western blot analysis (Fig. a) was performed using Modified whole cell extracts (1% SDS) from the HDAC1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-HDAC1 Polyclonal Antibody (Product # PA1-41120, 2µg/ml) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Densitometric analysis of this Western Blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to HDAC1..

HDAC1 Antibody in Western Blot (WB) — Western blot analysis of HDAC1 in 0.5 mg/mL Hek293 lysate. Samples were incubated in HDAC1 polyclonal antibody (Product # PA1-41120). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

HDAC1 Antibody in Western Blot (WB) — Western blot analysis of HDAC1 in HEK293 cell lysate. Sample was incubated in HDAC1 polyclonal antibody (Product # PA1-41120).

HDAC1 Antibody in ChIP Assay (ChIP) — Chromatin Immunoprecipitation (ChIP) assay of endogenous HDAC1 protein using HDAC1 Polyclonal Antibody: ChIP was performed using HDAC1 Polyclonal Antibody (Product # PA1-41120, 5 µg) on sheared chromatin from MCF-7 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to CDKN1-PR, GAPDH- TSS and FOSL1-PR as active binding regions and FOSL1- Exon1 and SAT alpha as inactive binding regions. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method. PR: Promoter, TSS: Transcription start site.

HDAC1 Antibody in ChIP Assay (ChIP) — Chromatin Immunoprecipitation (ChIP) assay of endogenous HDAC1 protein using Anti-HDAC1 Antibody: ChIP was performed using Anti-HDAC1 Rabbit Polyclonal Antibody (Product # PA1-41120, 5 µg) on sheared chromatin from HeLa cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to CDKN1A Intron 1, transcriptional start site and gene body (+2kb) of GAPDH, RPL30 exon 5 and SATA satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

HDAC1 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein in HDAC1 KO cell line compared to control cell line using Anti-HDAC1 Polyclonal Antibody (Product # PA1-41120). {KO}

HDAC1 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. Knockdown of HDAC1 was achieved by transfecting MCF-7 cells with HDAC1 specific siRNAs (Silencer® select Product # S73 and S75). ChIP was performed using HDAC1 Polyclonal Antibody (Product # PA1-41120) on sheared chromatin from MCF-7 knockdown cells (grey bar), non-specific scrambled siRNA transfected cells (orange bar) and untransfected cells (blue bar). Decrease in fold enrichment of active binding region in siRNA mediated knockdown cells confirms that antibody is specific to HDAC1. {KD}

HDAC1 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. MCF7 cells were transfected with HDAC1 siRNA and decrease in signal intensity was observed in western blot application using Anti- HDAC1 Polyclonal Antibody (Product # PA1-41120). {KD}

Product Details

PA1-41120

Applications	Tested Dilution	Publications
Western Blot (WB)	2 µg/mL	-
Immunohistochemistry (Paraffin) (IHC (P))	1:10-1:500	-
Immunohistochemistry (Frozen) (IHC (F))	1:10-1:500	-
Immunocytochemistry (ICC/IF)	1 µg/mL	-
ChIP assay (ChIP)	2.5 µg/10^6 cells	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Synthetic peptides corresponding to residues 1-5, 433-448, and 467-482 of human HDaC1. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein G
Storage buffer	PBS
Contains	0.05% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_1074972

Product Specific Information

Suggested positive control: 293 whole cell lysate, antigen standard for HDAC1 (transient overexpression lysate).

Target Information

Histone acetylation and deacetylation, catalyzed by multisubunit complexes, play a key role in the regulation of eukaryotic gene expression. HDAC1 belongs to the histone deacetylase/acuc/apha family and is a component of the histone deacetylase complex. It also interacts with retinoblastoma tumor-suppressor protein and this complex is a key element in the control of cell proliferation and differentiation. Together with metastasis-associated protein-2, HDAC1 deacetylates p53 and modulates its effect on cell growth and apoptosis.

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

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Bioinformatics

Protein Aliases: DKFZp686H12203; HD1; Histone deacetylase 1; Protein deacetylase HDAC1; Protein decrotonylase HDAC1; reduced potassium dependency, yeast homolog-like 1; RP4-811H24.2

Gene Aliases: GON-10; HD1; HDAC1; Hdac1-ps; MommeD5; RPD3; RPD3L1

UniProt ID: (Human) Q13547, (Mouse) O09106, (Rat) Q4QQW4

Entrez Gene ID: (Human) 3065, (Mouse) 433759, (Rat) 297893

negative regulation of transcription from RNA polymerase II promoter response to amphetamine transcription, DNA-templated regulation of transcription, DNA-templated transcription from RNA polymerase II promoter protein deacetylation endoderm development circadian rhythm positive regulation of cell proliferation negative regulation of cell proliferation epidermal cell differentiation negative regulation of gene expression positive regulation of receptor biosynthetic process chromatin modification histone deacetylation hippocampus development neuron differentiation positive regulation of interleukin-1 production positive regulation of tumor necrosis factor production circadian regulation of gene expression cellular response to oxidative stress odontogenesis of dentin-containing tooth positive regulation of tyrosine phosphorylation of Stat3 protein embryonic digit morphogenesis ATP-dependent chromatin remodeling negative regulation of apoptotic process negative regulation of I-kappaB kinase/NF-kappaB signaling negative regulation of neuron apoptotic process negative regulation by host of viral transcription negative regulation of transcription, DNA-templated positive regulation of transcription, DNA-templated positive regulation of transcription from RNA polymerase II promoter negative regulation of insulin secretion rhythmic process positive regulation of oligodendrocyte differentiation negative regulation of androgen receptor signaling pathway hair follicle placode formation eyelid development in camera-type eye fungiform papilla formation histone H3 deacetylation histone H4 deacetylation negative regulation of canonical Wnt signaling pathway positive regulation of chemokine (C-X-C motif) ligand 2 production positive regulation of type B pancreatic cell apoptotic process negative regulation of peptidyl-lysine acetylation negative regulation of intrinsic apoptotic signaling pathway chromatin remodeling methylation-dependent chromatin silencing blood coagulation negative regulation of myotube differentiation viral process regulation of signal transduction by p53 class mediator beta-catenin-TCF complex assembly response to caffeine response to lipopolysaccharide response to drug response to hyperoxia cellular response to tumor necrosis factor

Process(es)

transcription regulatory region sequence-specific DNA binding 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 transcription factor binding RNA polymerase II repressing transcription factor binding RNA polymerase II transcription corepressor activity transcription factor activity, sequence-specific DNA binding histone deacetylase activity protein binding transcription factor binding deacetylase activity enzyme binding nucleosomal DNA binding NAD-dependent histone deacetylase activity (H3-K14 specific) protein deacetylase activity activating transcription factor binding histone deacetylase binding transcription regulatory region DNA binding protein N-terminus binding NF-kappaB binding repressing transcription factor binding DNA binding chromatin binding transcription corepressor activity hydrolase activity histone deacetylase activity (H3-K14 specific) histone deacetylase activity (H3-K9 specific) protein complex binding 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)