Recombinant Anti-mSin3a antibody [EPR27103-53] (ab307197)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR27103-53] to mSin3A
- Suitable for: Flow Cyt (Intra), WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-mSin3a antibody [EPR27103-53]
See all mSin3A primary antibodies -
Description
Rabbit monoclonal [EPR27103-53] to mSin3A -
Host species
Rabbit -
Specificity
IP: not suitable with mouse.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IPmore details
Unsuitable for: ChIP,ICC/IF or IHC-P -
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: Wild-type HAP1 whole cell lysate. HeLa, K562, 293T, C6, RAW264.7 and F9 whole cell lysate. Human and mouse testis tissue lysate. Flow Cyt (Intra): Wild-type HAP1 cells. F9 cells. IP: HAP1 whole cell lysate.
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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 +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR27103-53 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab307197 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) |
1/500.
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WB |
1/1000. Detects a band of approximately 170 kDa.
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Detects a band of approximately 170 kDa. |
IP
1/30. |
Target
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Function
Acts as a transcriptional repressor. Interacts with MXI1 to repress MYC responsive genes and antagonize MYC oncogenic activities. Also interacts with MAD-MAX heterodimers by binding to MAD. The heterodimer then represses transcription by tethering SIN3A to DNA. Acts as a corepressor for REST. -
Sequence similarities
Contains 3 PAH (paired amphipathic helix) domains. -
Post-translational
modificationsSUMO1 sumoylated by TOPORS. Probably desumoylated by SENP2. -
Cellular localization
Nucleus. Nucleus > nucleolus. Recruited to the nucleolus by SAP30L. - Information by UniProt
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Database links
- Entrez Gene: 25942 Human
- Entrez Gene: 20466 Mouse
- Entrez Gene: 363067 Rat
- Omim: 607776 Human
- SwissProt: Q96ST3 Human
- SwissProt: Q60520 Mouse
- Unigene: 513039 Human
- Unigene: 15755 Mouse
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Alternative names
- AW553200 antibody
- DKFZP434K2235 antibody
- FLJ90319 antibody
see all
Images
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All lanes : Anti-mSin3a antibody [EPR27103-53] (ab307197) at 1/1000 dilution
Lane 1 : Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate
Lane 2 : mSin3A knockout HAP1 whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) at 1/10000 dilutionBlocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
Lysates at 20 µg per lane.
The samples were run on a Bis-Tris gel.
Performed under reducing conditions.
False colour image of Western blot: Anti-mSin3A antibody (ab307197) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab307197 was shown to bind specifically to mSin3A. A band was observed at 170 kDa in wild-type HAP1 cell lysates with no signal observed at this size in mSin3A knockout cell line. To generate this image, wild-type and mSin3A knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-mSin3a antibody [EPR27103-53] (ab307197) at 1/1000 dilution
Lane 1 : Human testis tissue lysate
Lane 2 : Mouse testis tissue lysate
Lane 3 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 4 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 5 : F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilutionBlocking and diluting buffer and concentration: 5% NFDM/TBST.
The identity of the bands below 75 kDa are unknown.
The blot was developed using a higher sensitivity ECL substrate.
Exposure time: 180 seconds.
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All lanes : Anti-mSin3a antibody [EPR27103-53] (ab307197) at 1/1000 dilution
Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate 20 µg
Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilutionBlocking and diluting buffer and concentration: 5% NFDM/TBST.
The identity of the bands below 75 kDa are unknown.
This blot was developed using a high sensitivity ECL substrate.
Exposure time: 180 seconds.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) (Right panel) compared to mSin3A knockout HAP1 (Left panel) cells labeling mSin3A with AB307197 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).
A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labeling mSin3A with AB307197 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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mSin3A was immunoprecipitated from 0.35 mg HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate 10 µg with AB307197 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using AB307197 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HAP1 whole cell lysate 10 µg
Lane 2: AB307197 IP in HAP1 whole cell lysate 10 ug
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab307197 in HAP1 cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 6 seconds.
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
Certificate of Compliance
References (0)
ab307197 has not yet been referenced specifically in any publications.