Anti-WHSC1/NSD2 antibody [29D1] - BSA and Azide free (ab235726)
Key features and details
- Mouse monoclonal [29D1] to WHSC1/NSD2 - BSA and Azide free
- Suitable for: WB, IP, IHC-P
- Reacts with: Human
- Isotype: IgG2b
Overview
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Product name
Anti-WHSC1/NSD2 antibody [29D1] - BSA and Azide free
See all WHSC1/NSD2 primary antibodies -
Description
Mouse monoclonal [29D1] to WHSC1/NSD2 - BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Fusion protein corresponding to Human WHSC1/NSD2 aa 1-650.
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Positive control
- IHC-P: Human normal testis tissue. WB: HeLa and SHSY5Y whole cell lysates. IP: HepG2 cell lysate.
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General notes
ab235726 is a PBS only version of ab75359.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
29D1 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
Images
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All lanes : Anti-WHSC1/NSD2 antibody [29D1] (ab75359)
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : WHSC1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 152 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab75359).
Lanes 1- 2: Merged signal (red and green). Green - ab75359 observed at 80 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.
ab75359 was shown to react with WHSC1/NSD2 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264936 (knockout cell lysate ab257798) lane below 80kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and WHSC1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab75359 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at a 5 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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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 data was developed using the same antibody clone in a different buffer formulation containing PBS, Azide and Arginine (ab75359).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, Azide and Arginine (ab75359).