Overview

  • Product name

    Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade
    See all WHSC1/NSD2 primary antibodies
  • Description

    Mouse monoclonal [29D1] to WHSC1/NSD2 - ChIP Grade
  • Host species

    Mouse
  • Tested applications

    Suitable for: CHIPseq, WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Fusion protein corresponding to Human WHSC1/NSD2 aa 1-650.

  • Positive control

    • IHC-P: Human normal testis tissue. WB: HeLa and SHSY5Y whole cell lysates. IP: HepG2 cell lysate.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Function

    Probable histone methyltransferase (By similarity). May act as a transcription regulator that binds DNA and suppresses IL5 transcription.
  • Tissue specificity

    Widely expressed.
  • Involvement in disease

    Note=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 similarities

    Belongs 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.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Cytoplasm and Nucleus. Chromosome.
  • Information by UniProt
  • Database links

  • Alternative names

    • FLJ23286 antibody
    • IL5 promoter REII region binding protein antibody
    • KIAA1090 antibody
    • MGC176638 antibody
    • MMSET antibody
    • MMSET type II antibody
    • Multiple myeloma SET domain containing protein type III antibody
    • Multiple myeloma SET domain protein antibody
    • Multiple myeloma SET domain-containing protein antibody
    • NSD 2 antibody
    • NSD2 antibody
    • NSD2_HUMAN antibody
    • Nuclear receptor binding SET domain protein 2 antibody
    • Nuclear SET domain-containing protein 2 antibody
    • Probable histone-lysine N-methyltransferase NSD2 antibody
    • Protein trithorax-5 antibody
    • REIIBP antibody
    • Trithorax/ash1 related protein 5 antibody
    • TRX5 antibody
    • TRX5 protein antibody
    • WHS antibody
    • Whsc1 antibody
    • Wolf Hirschhorn syndrome candidate 1 antibody
    • Wolf Hirschhorn syndrome candidate 1 protein antibody
    • Wolf-Hirschhorn syndrome candidate 1 protein antibody
    see all

Images

  • 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.

    Secondary
    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.

    See Abreview

  • 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.

    Secondary
    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. 

  • 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.

References

This product has been referenced in:

  • Wang JJ  et al. Histone methyltransferase NSD2 mediates the survival and invasion of triple-negative breast cancer cells via stimulating ADAM9-EGFR-AKT signaling. Acta Pharmacol Sin N/A:N/A (2019). Read more (PubMed: 30670815) »
  • Mills A  et al. Wolf-Hirschhorn Syndrome-Associated Genes Are Enriched in Motile Neural Crest Cells and Affect Craniofacial Development in Xenopus laevis. Front Physiol 10:431 (2019). Read more (PubMed: 31031646) »
See all 13 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (SH-SY5Y Neuroblastoma Cell Line)
Permeabilization
Yes - 0.2% Triton-X100 for 5 min
Specification
SH-SY5Y Neuroblastoma Cell Line
Blocking step
10% Normal Donkey Serum + 0.3M Glycin in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Paraformaldehyde

Dr. Lars Riecken

Verified customer

Submitted Oct 26 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (SH-SY5Y Neuroblastoma Cell Line)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
15 µg
Specification
SH-SY5Y Neuroblastoma Cell Line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Oct 24 2018

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (BaF3 cell line NIH3T3 cell line HEK 293 cell line)
Specification
BaF3 cell line NIH3T3 cell line HEK 293 cell line
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 01 2013

Answer

Thank you for contacting us.

Please find attached the codes;

XXXXXXX
XXXXXXX
XXXXXXX


We do not provide codes for linking antibodies to different molecules.

Please check the terms and conditions by clicking the following link;

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (hct116, hela cells)
Loading amount
50 µg
Specification
hct116, hela cells
Gel Running Conditions
Reduced Denaturing (10% tris glycine)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Apr 30 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up