Product nameAnti-WHSC1/NSD2 antibody [29D1] - ChIP Grade
See all WHSC1/NSD2 primary antibodies
DescriptionMouse monoclonal [29D1] to WHSC1/NSD2 - ChIP Grade
Tested applicationsSuitable for: CHIPseq, WB, IP, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Fusion protein, corresponding to amino acids 1-647 of Human GST-WHSC1/NSD2
- IHC-P: Human normal testis tissue. WB: HeLa and SHSY5Y whole cell lysates. IP: HepG2 cell lysate.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Some batches contain 0.4M arginine. Please contact Scientific Support for further information.
Concentration information loading...
PurityProtein G purified
Light chain typekappa
ChIP Related Products
Our Abpromise guarantee covers the use of ab75359 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|CHIPseq||Use at an assay dependent concentration. PubMed: 22196736|
|WB||Use a concentration of 5 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 152 kDa).|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionProbable histone methyltransferase (By similarity). May act as a transcription regulator that binds DNA and suppresses IL5 transcription.
Tissue specificityWidely expressed.
Involvement in diseaseNote=A chromosomal aberration involving WHSC1 is a cause of multiple myeloma tumors. Translocation t(4;14)(p16.3;q32.3) with IgH.
Note=WHSC1 is located in the Wolf-Hirschhorn syndrome (WHS) critical region. WHS results from by sub-telomeric deletions in the short arm of chromosome 4. WHSC1 is deleted in every case, however deletion of linked genes contributes to both the severity of the core characteristics and the presence of the additional syndromic problems.
Sequence similaritiesBelongs to the histone-lysine methyltransferase family. SET2 subfamily.
Contains 1 AWS domain.
Contains 1 HMG box DNA-binding domain.
Contains 4 PHD-type zinc fingers.
Contains 1 post-SET domain.
Contains 2 PWWP domains.
Contains 1 SET domain.
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationCytoplasm and Nucleus. Chromosome.
- Information by UniProt
- FLJ23286 antibody
- IL5 promoter REII region binding protein antibody
- KIAA1090 antibody
ab75359 (1µg/ml) staining WHSC in human testis using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1/ 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.
All lanes : Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade (ab75359) at 1/5000 dilution
Lane 1 : Whole cell lysate prepared from HeLa cells
Lane 2 : Whole cell lysate prepared from HCT116 cells
Lysates/proteins at 50 µg per lane.
All lanes : HRP conjugated goat anti-mouse polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 152 kDa
Exposure time: 3 minutes
Blocked with 5% milk for 1 hour at 23°C.
All lanes : Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade (ab75359) at 5 µg/ml
All lanes : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 152 kDa
Observed band size: 180,80 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutes
The full-length MMSET-II protein has an apparent molecular weight of approximately 180 kDa, whereas the 645 amino acid long MMSET-I exhibits a mobility of approximately 80 kDa.
Abcam recommends blocking with 1% milk.
IHC image of WHSC1/NSD2 staining in Human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75359, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
WHSC1/NSD2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 10µg of Mouse monoclonal to WHSC1/NSD2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab75359.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 71kDa: WHSC1/NSD2.
This product has been referenced in:
- Li N et al. AKT-mediated stabilization of histone methyltransferase WHSC1 promotes prostate cancer metastasis. J Clin Invest 127:1284-1302 (2017). Read more (PubMed: 28319045) »
- Hernando H et al. EZH2 Inhibition Blocks Multiple Myeloma Cell Growth through Upregulation of Epithelial Tumor Suppressor Genes. Mol Cancer Ther 15:287-98 (2016). Read more (PubMed: 26590165) »