Recombinant Anti-HDAC2 antibody [Y461] (ab32117)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y461] to HDAC2
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seq
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-HDAC2 antibody [Y461]
See all HDAC2 primary antibodies -
Description
Rabbit monoclonal [Y461] to HDAC2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seqmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, A431, Hela and K562 cell lysate and rat brain tissue homogenate. IHC-P: Human breast carcinoma and rat spinal cord tissue. ICC/IF: MCF-7 and wildtype HAP1 cells. Flow Cyt (intra): HeLa cells. ChIC/CUT&RUN-Seq: K-562 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y461 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- HRP Anti-HDAC2 antibody [Y461] (ab195851)
- Alexa Fluor® 488 Anti-HDAC2 antibody [Y461] (ab196471)
- Alexa Fluor® 647 Anti-HDAC2 antibody [Y461] (ab196518)
- PE Anti-HDAC2 antibody [Y461] (ab210827)
- Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
- Alexa Fluor® 568 Anti-HDAC2 antibody [Y461] (ab214384)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32117 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (9) |
1/2000. Predicted molecular weight: 55 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Purified ab32117 at 1:1500 dilution. |
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ICC/IF |
1/250 - 1/500.
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IP |
1/60.
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Flow Cyt (Intra) |
1/60 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5 µg |
Notes |
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WB
1/2000. Predicted molecular weight: 55 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Purified ab32117 at 1:1500 dilution. |
ICC/IF
1/250 - 1/500. |
IP
1/60. |
Flow Cyt (Intra)
1/60 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5 µg |
Target
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Function
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. -
Tissue specificity
Widely expressed; lower levels in brain and lung. -
Sequence similarities
Belongs to the histone deacetylase family. HD type 1 subfamily. -
Post-translational
modificationsS-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. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 3066 Human
- Entrez Gene: 15182 Mouse
- Entrez Gene: 84577 Rat
- Omim: 605164 Human
- SwissProt: Q92769 Human
- SwissProt: P70288 Mouse
- Unigene: 3352 Human
- Unigene: 19806 Mouse
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Alternative names
- D10Wsu179e antibody
- HD 2 antibody
- HD2 antibody
see all
Images
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All lanes : Anti-HDAC2 antibody [Y461] (ab32117) at 1/1000 dilution
Lane 1 : HT-22 (mouse hippocampal neuronal cell) whole cell lysate
Lane 2 : SW10 (mouse neuronal Schwann cell) whole cell lysate
Lane 3 : bEnd.3 (mouse brain endothelioma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 26 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
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All lanes : Anti-HDAC2 antibody [Y461] (ab32117) at 1/1000 dilution
Lane 1 : GH3 (rat pituitary epithelial cell) whole cell lysate
Lane 2 : L6 (rat skeletal muscle myoblast) whole cell lysate
Lane 3 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 4 : AR42J (rat pancreatic tumor epithelial cell) whole cell lysate
Lane 5 : 2.4G2 (rat B cell lymphoma B lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 37 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)
Immunohistochemical analysis of paraffin-embedded fixed (A) Wild-type HEK293T (human embryonic kidney epithelial cell) cell pellet. (B) HDAC2 knockout HEK293T (ab266590) cell pellet staining HDAC2 with ab32117 at 1/10000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counter-staining used was hematoxylin.
Positive staining on (A) Wild-type HEK293T cell pellet, no staining on HDAC2 knockout HEK293T (ab266590) cell pellet.
The section was incubated with ab32117 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrumentHeat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunocytochemistry/ Immunofluorescence analysis of Wild-type HEK293T/HDAC2 KO HEK293T (HDAC2 knockout human embryonic kidney epithelial cell) (ab266590) cells labeling HDAC2 with ab32117 at 1/200 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 dilution (2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (2.5 µg/ml). DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Confocal image showing nuclear staining in Parental HEK293T cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2 x 10^5 K-562 (Human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab32117 [Y461] . The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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All lanes : Anti-HDAC2 antibody [Y461] (ab32117) at 1/10000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : Mouse brain lysate
Lane 3 : Rat brain lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 55 kDa -
Purified ab32117 at 1:20 dilution (0.5µg) immunoprecipitating HDAC2 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg.
Lane 2 (+): ab32117 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32127 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP)(ab131366) (1:5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: kDa -
Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labelling HDAC2 with Purified ab32117 at 1:20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HDAC2 with Purified ab32117 at 1:50 dilution (2.1 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue sections labeling HDAC2 with Purified ab32117 at 1:1500 dilution (0.071 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling HDAC2 with Purified ab32117 at 1:1500 dilution (0.071 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue sections labeling HDAC2 with Purified ab32117 at 1:1500 dilution (0.071 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling HDAC2 with Purified ab32117 at 1:1500 dilution (0.071 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: HDAC2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: A431 whole cell lysate (20 µg)
Lane 4: Hela whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab32117 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32117 was shown to specifically react with HDAC2 when HDAC2 knockout samples were used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab32117 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 and 1/10000 respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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ab32117 staining HDAC2 in wild-type HAP1 cells (top panel) and HDAC2 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32117 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (83)
ab32117 has been referenced in 83 publications.
- Wu K et al. Green tea polyphenols inhibit malignant melanoma progression via regulating circ_MITF/miR-30e-3p/HDAC2 axis. Biotechnol Appl Biochem 69:808-821 (2022). PubMed: 33797132
- Chen KQ et al. Quanzhenyiqitang Reverses LPS-Induced Inflammation via Inhibiting PYK2/p38MAPK/HDAC2/CK2 Signaling Pathway in Rat Alveolar Macrophage. Evid Based Complement Alternat Med 2022:7857022 (2022). PubMed: 35047050
- Yuan B et al. Patient-derived organoids for personalized gallbladder cancer modelling and drug screening. Clin Transl Med 12:e678 (2022). PubMed: 35075805
- Lu Z et al. Promotion of microRNA-146a by histone deacetylase 4 silencing contributes to radiosensitization of esophageal carcinoma. J Transl Med 20:101 (2022). PubMed: 35193602
- Ma F et al. MicroRNA-455-3p functions as a tumor suppressor by targeting HDAC2 to regulate cell cycle in hepatocellular carcinoma. Environ Toxicol 37:1675-1685 (2022). PubMed: 35286011