Anti-WHSC1/NSD2 antibody [29D1]

• WB

Lab

Western blot - Anti-WHSC1/NSD2 antibody [29D1] (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.

All lanes:

Western blot - Anti-WHSC1/NSD2 antibody [29D1] (ab75359) at 5 µg/mL

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human WHSC1 (NSD2) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-whsc1-nsd2-knockout-hela-cell-lysate-ab257798'>ab257798</a>) at 20 µg

Predicted band size: 152 kDa

Observed band size: 80 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-WHSC1/NSD2 antibody [29D1] (AB75359)

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 70^oC; 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:

Immunoprecipitation - Anti-WHSC1/NSD2 antibody [29D1] (ab75359)

Predicted band size: 152 kDa

false

• WB

AbReview29576****

Western blot - Anti-WHSC1/NSD2 antibody [29D1] (AB75359)

Blocked with 5% milk for 1 hour at 23°C.

All lanes:

Western blot - Anti-WHSC1/NSD2 antibody [29D1] (ab75359) at 1/5000 dilution

Lane 1:

Whole cell lysate prepared from HeLa cells at 50 µg

Lane 2:

Whole cell lysate prepared from HCT116 cells at 50 µg

Secondary

All lanes:

HRP conjugated goat anti-mouse polyclonal at 1/10000 dilution

Predicted band size: 152 kDa

true

Exposure time: 3min

Image courtesy of an anonymous Abreview.

• WB

Lab

Western blot - Anti-WHSC1/NSD2 antibody [29D1] (AB75359)

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.

All lanes:

Western blot - Anti-WHSC1/NSD2 antibody [29D1] (ab75359) at 5 µg/mL

All lanes:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 152 kDa

Observed band size: 180 kDa,80 kDa

false

Exposure time: 20min