Recombinant Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR9913] to MBD3 - ChIP Grade
- Suitable for: Flow Cyt (Intra), IP, WB, ICC/IF, ChIP
- Knockout validated
- Reacts with: Mouse, Rat, Cow, Human
Related conjugates and formulations
Overview
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Product name
Anti-MBD3 antibody [EPR9913] - ChIP Grade
See all MBD3 primary antibodies -
Description
Rabbit monoclonal [EPR9913] to MBD3 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, WB, ICC/IF, ChIPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HeLa, 293T, fetal brain and Y79 lysates; HeLa cells; Permeabilized 293T 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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20ºC. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR9913 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab157464 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) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IP | (1) |
Use at an assay dependent concentration.
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WB | (4) |
1/1000 - 1/5000. Predicted molecular weight: 33 kDa.
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ICC/IF | (2) |
1/100 - 1/1000.
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ChIP | (1) |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IP
Use at an assay dependent concentration. |
WB
1/1000 - 1/5000. Predicted molecular weight: 33 kDa. |
ICC/IF
1/100 - 1/1000. |
ChIP
Use at an assay dependent concentration. |
Target
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Function
Does not bind DNA by itself. Recruits histone deacetylases and DNA methyltransferases. Acts as transcriptional repressor and plays a role in gene silencing. -
Sequence similarities
Contains 1 MBD (methyl-CpG-binding) domain. -
Cellular localization
Nucleus. Nuclear, in discrete foci. - Information by UniProt
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Database links
- Entrez Gene: 616090 Cow
- Entrez Gene: 53615 Human
- Entrez Gene: 17192 Mouse
- Entrez Gene: 362834 Rat
- Omim: 603573 Human
- SwissProt: O95983 Human
- SwissProt: Q9Z2D8 Mouse
- Unigene: 178728 Human
see all -
Alternative names
- AI181826 antibody
- AU019209 antibody
- MBD 3 antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ProteinX knockout HAP1 cell lysate (20 µg)
Lane 3: Wild-type HAP1 cell lysate (20 µg)
Lane 4: ProteinX knockout HAP1 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab157464 observed at 33 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab157464 was shown to recognize MBD3 when MBD3 knockout samples were used, along with additional cross-reactive bands. Wild-type and MBD3 knockout samples were subjected to SDS-PAGE. ab157464 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging. -
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling MBD3 with purified ab157464 at 1/1000. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
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Chromatin was prepared from HeLa cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab157464 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
All lanes : Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysate
Lane 3 : Fetal brain lysate
Lane 4 : Y79 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 33 kDa -
Chromatin was prepared from NIH/3T3 cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab157464 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
Immunofluorescent analysis of HeLa cells labeling MBD3 with ab157464 at 1/100 dilution.
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Intracellular flow cytometric analysis of permeabilized 293T cells labeling MBD3 with ab157464 at 1/10 dilution (red) compared to a rabbit IgG negative control (green).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (17)
ab157464 has been referenced in 17 publications.
- Xu H et al. Choroid plexus NKCC1 mediates cerebrospinal fluid clearance during mouse early postnatal development. Nat Commun 12:447 (2021). PubMed: 33469018
- Ishibashi T et al. Methyl-CpG binding domain protein 3: a new diagnostic marker and potential therapeutic target of melanoma. Biosci Trends 14:390-395 (2020). PubMed: 32963182
- Low JKK et al. The Nucleosome Remodeling and Deacetylase Complex Has an Asymmetric, Dynamic, and Modular Architecture. Cell Rep 33:108450 (2020). PubMed: 33264611
- Ragheb R et al. Differential regulation of lineage commitment in human and mouse primed pluripotent stem cells by the nucleosome remodelling and deacetylation complex. Stem Cell Res 46:101867 (2020). PubMed: 32535494
- Kong X et al. MTA1, a Target of Resveratrol, Promotes Epithelial-Mesenchymal Transition of Endometriosis via ZEB2. Mol Ther Methods Clin Dev 19:295-306 (2020). PubMed: 33145366