Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade

Name	Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade
Supplier	Abcam
Catalog	ab75359
Prices	$400.00
Sizes	100 µg
Host	Mouse
Clonality	Monoclonal
Isotype	IgG2b
Clone	29D1
Applications	ChIPseq WB IP IHC-P
Species Reactivities	Mouse, Human
Antigen	Fusion protein, corresponding to amino acids 1-647 of Human GST-WHSC1/NSD2
Description	Mouse Monoclonal
Gene	WHSC1
Conjugate	Unconjugated
Supplier Page	Shop

Product images

All lanes : Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade (ab75359) at 5 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutionPerformed under reducing conditions.

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.

All lanes : Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade (ab75359) at 1/5000 dilutionLane 1 : Whole cell lysate prepared from HeLa cellsLane 2 : Whole cell lysate prepared from HCT116 cellsLysates/proteins at 50 µg per lane.SecondaryHRP conjugated goat anti-mouse polyclonal at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade (ab75359) at 1/700 dilution + HeLa nuclear lysate at 20 µgSecondaryHRP-conjugated goat anti-mouse IgG2b polyclonal at 1/1000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

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.

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