Anti-MDMX/MDM4 antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MDMX/MDM4 antibody (AB16058)

ab16058 staining MDMX/MDM4 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab16058 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Unknown

Western blot - Anti-MDMX/MDM4 antibody (AB16058)

Two bands were detected with the antibody in HeLa nuclear extract, at ~80 and 100 kDa. The 80 kDa band was no longer detected after the addition of competing peptide, suggesting that this band corresponds to MDMX/MDM4. The 100 kDa band is not competed away by the addition of the immunizing peptide, suggesting that it is nonspecific cross-reactivity.

All lanes:

Western blot - Anti-MDMX/MDM4 antibody (ab16058) at 1/1000 dilution

Lane 1:

20ug calf thymus histone

Lane 2:

20ug HeLa nuclear extract

Lane 3:

20ug calf thymus histone with Human MDMX/MDM4 peptide (<a href='/en-us/products/unavailable/human-mdmxmdm4-peptide-ab16383'>ab16383</a>)

Lane 4:

20ug HeLa nuclear extract with Human MDMX/MDM4 peptide (<a href='/en-us/products/unavailable/human-mdmxmdm4-peptide-ab16383'>ab16383</a>)

Secondary

All lanes:

Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/5000 dilution

Predicted band size: 55 kDa

Observed band size: 100 kDa,80 kDa

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