Recombinant Anti-HDAC2 antibody [EPR20117] - BSA and Azide free (ab251561)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20117] to HDAC2 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IP, ICC/IF, IHC-P, ChIC/CUT&RUN-seq, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-HDAC2 antibody [EPR20117] - BSA and Azide free
See all HDAC2 primary antibodies -
Description
Rabbit monoclonal [EPR20117] to HDAC2 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, ICC/IF, IHC-P, ChIC/CUT&RUN-seq, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: His-tagged human HDAC2 recombinant protein (aa339-488);HeLa,SH-SY5Y,HEK-293,PC-12,NIH/3T3 whole cell lysates; Human fetal brain,fetal heart and fetal kidney lysates; Mouse brain and heart lysates; Rat heart,brain and spleen lysates IHC-P: Human testis,tonsil,prostate hyperplasia,prostate cancer,breast cancer and synovial sarcoma tissues; mouse colon tissue and rat spleen tissue ICC/IF: HEK-293 and NIH/3T3 cells Flow Cyt (intra): NIH/3T3 cells IP: HeLa cell lysate ChIC/CUT&RUN-Seq: K-562 cells
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General notes
ab251561 is the carrier-free version of ab219053.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20117 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab251561 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). |
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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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 K-562 (Human chronic myelogenous leukemia lymphoblast) cells and 5µg of ab219053 [EPR20117]. 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.
This data was developed using the same antibody clone in a different buffer formulation (ab219053).
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All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HDAC2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab219053).
Lanes 1- 2: Merged signal (red and green). Green - ab219053 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab219053 was shown to react with HDAC2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266589 (knockout cell lysate ab256938) was used. Wild-type HEK-293T and HDAC2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab219053 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HDAC2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab219053).
Lanes 1-2: Merged signal (red and green). Green - ab219053 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.
ab219053 Anti-HDAC2 antibody [EPR20117] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HDAC2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab219053).
Lanes 1-2: Merged signal (red and green). Green - ab219053 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.
ab219053 Anti-HDAC2 antibody [EPR20117] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266588 (knockout cell lysate ab256937) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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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Datasheet download
Certificate of Compliance
References (0)
ab251561 has not yet been referenced specifically in any publications.