Anti-HIPK2 antibody [EPR3819] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HIPK2 antibody [EPR3819] - BSA and Azide free (AB221980)

This data was developed using ab108543, the same antibody clone in a different buffer formulation.

Overlay histogram showing HEK293 cells stained with ab108543 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab108543, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• WB

Unknown

Western blot - Anti-HIPK2 antibody [EPR3819] - BSA and Azide free (AB221980)

This data was developed using ab108543, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-HIPK2 antibody [EPR3819] (<a href='/en-us/products/primary-antibodies/hipk2-antibody-epr3819-ab108543'>ab108543</a>) at 1/2000 dilution

Lane 1:

HT-1080 cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 131 kDa

Observed band size: 100 kDa,130 kDa

false

• WB

Lab

Western blot - Anti-HIPK2 antibody [EPR3819] - BSA and Azide free (AB221980)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This data was developed using ab108543, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-HIPK2 antibody [EPR3819] (<a href='/en-us/products/primary-antibodies/hipk2-antibody-epr3819-ab108543'>ab108543</a>) at 1/2000 dilution

All lanes:

Mouse heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 131 kDa

Observed band size: 100 kDa,130 kDa

false

Exposure time: 180s