Anti-mSin3A antibody - ChIP Grade (ab3479)
Key features and details
- Rabbit polyclonal to mSin3A - ChIP Grade
- Suitable for: ChIP, ICC/IF, WB, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
-
Product name
Anti-mSin3A antibody - ChIP Grade
See all mSin3A primary antibodies -
Description
Rabbit polyclonal to mSin3A - ChIP Grade -
Host species
Rabbit -
Tested Applications & Species
Application Species ChIP HumanICC/IF MouseHumanIHC-P HumanWB Human -
Immunogen
Synthetic peptide corresponding to Mouse mSin3A aa 1-19.
Sequence:MKRRLDDQESDVYAAQQRR
(Peptide available asab4995) -
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, PBS -
Concentration information loading...
-
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
-
ChIP Related Products
-
Compatible Secondaries
-
Immunizing Peptide (Blocking)
-
Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab3479 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
---|---|
ChIP |
Human
|
ICC/IF |
Mouse
Human
|
IHC-P |
Human
|
WB |
Human
|
Application | Abreviews | Notes |
---|---|---|
ChIP | (2) |
Use at an assay dependent concentration.
|
ICC/IF | (1) |
1/50 - 1/500.
|
WB | (5) |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 150 kDa (predicted molecular weight: 145 kDa).Can be blocked with mSin3A peptide (ab4995).
|
IHC-P | (1) |
Use a concentration of 2 µg/ml.
|
Notes |
---|
ChIP
Use at an assay dependent concentration. |
ICC/IF
1/50 - 1/500. |
WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 150 kDa (predicted molecular weight: 145 kDa).Can be blocked with mSin3A peptide (ab4995). |
IHC-P
Use a concentration of 2 µg/ml. |
Target
-
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
-
Database links
- Entrez Gene: 25942 Human
- Entrez Gene: 20466 Mouse
- Omim: 607776 Human
- SwissProt: Q96ST3 Human
- SwissProt: Q60520 Mouse
- Unigene: 513039 Human
- Unigene: 15755 Mouse
-
Alternative names
- AW553200 antibody
- DKFZP434K2235 antibody
- FLJ90319 antibody
see all
Images
-
Western blot of mSin3A on K562 cell extract using ab3479.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mSin3A antibody - ChIP Grade (ab3479)
Paraffin-embedded human ovary carcinoma tissue (right panel) stained for mSin3A using ab3479 at 1/200 dilution compared to a negative control without primary antibody (left panel) in immunohistochemical analysis, followed by HRP-conjugated secondary antibody. Detection: DAB staining.
Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min.
-
ChIP analysis of Sin3A was performed using cross-linked chromatin from 1x106 HCT 116 (human colorectal carcinoma cell line) cells treated with serum for 0, 15, and 30 minutes. IP was performed using a multiplex microplate Matrix ChIP assay with 1.0 µl/100 µl well volume of ab3479. Chromatin aliquots from ~1x105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µl of eluted DNA in 2 µl SYBR real-time PCR reactions containing primers to amplify -15kb upstream of the Egr1 gene or exon-1 or exon-2 of Egr1. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. A schematic representation of the Egr-1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars.
-
Immunocytochemical immunoflurescence analysis of NIH-3T3 cells labelling mSin3A using ab3479. Formalin-fixed cells were permeablized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were then incubated with ab3479 in 3% BSA-PBS at a dilution of 1:100 overnight in a 4°C high humidity environment. Cells were then washed with PBST and incubated with a green DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Cells were counterstained with DAPI or Hoechst labelling the nuclear DNA blue. The Left Image is a negative control without the prescence of ab3479. Image magnification is 60X.
-
Immunocytochemical immunoflurescence analysis of HeLa cells labelling mSin3A using ab3479. Formalin-fixed cells were permeablized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were then incubated with ab3479 in 3% BSA-PBS at a dilution of 1:200 overnight in a 4°C high humidity environment. Cells were then washed with PBST and incubated with a green DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Cells were counterstained blue with DAPI or Hoechst labelling the nuclear DNA and red against actin using an Alexa Fluor® 554 conjugate. The Left Image is a negative control without the prescence of ab3479. Image magnification is 60X.
-
Immunocytochemical immunoflurescence analysis of NIH-3T3 cells labelling mSin3A using ab3479. Formalin-fixed cells were permeablized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were then incubated with ab3479 in 3% BSA-PBS at a dilution of 1:100 overnight in a 4°C high humidity environment. Cells were then washed with PBST and incubated with a green DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Cells were counterstained blue with DAPI or Hoechst labelling the nuclear DNA and red against actin using an Alexa Fluor® 554 conjugate. The Left Image is a negative control without the prescence of ab3479. Image magnification is 60X.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mSin3A antibody - ChIP Grade (ab3479)ab3479 (2µg/ml) staining mSin3A in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of nuclei of epithelial cells.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1/ EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (36)
ab3479 has been referenced in 36 publications.
- Hwang I et al. CIC is a critical regulator of neuronal differentiation. JCI Insight 5:N/A (2020). PubMed: 32229723
- Pinto DO et al. Effect of transcription inhibition and generation of suppressive viral non-coding RNAs. Retrovirology 16:13 (2019). PubMed: 31036006
- Fuller NO et al. CoREST Complex-Selective Histone Deacetylase Inhibitors Show Prosynaptic Effects and an Improved Safety Profile To Enable Treatment of Synaptopathies. ACS Chem Neurosci 10:1729-1743 (2019). PubMed: 30496686
- Zhu J et al. NKX2-8 deletion-induced reprogramming of fatty acid metabolism confers chemoresistance in epithelial ovarian cancer. EBioMedicine 43:238-252 (2019). PubMed: 31047858
- Meier-Soelch J et al. RNAi-Based Identification of Gene-Specific Nuclear Cofactor Networks Regulating Interleukin-1 Target Genes. Front Immunol 9:775 (2018). PubMed: 29755455