Anti-Rsk 2 / MAPKAP Kinase 1b antibody [Y82] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rsk 2 / MAPKAP Kinase 1b antibody [Y82] - BSA and Azide free (AB247218)

This data was developed using ab32062, the same antibody clone in a different buffer formulation.

ab32062 at a 1 : 250 - 1 : 500 dilution staining Rsk 2 / MAPKAP Kinase 1b in human urinary bladder carcinoma, using Immunohistochemistry, Paraffin Embedded Tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Rsk 2 / MAPKAP Kinase 1b antibody [Y82] - BSA and Azide free (AB247218)

This data was developed using ab32062, the same antibody clone in a different buffer formulation.Overlay histogram showing MCF7 cells stained with ab32062 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab32062, 1/1000 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 MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• WB

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Western blot - Anti-Rsk 2 / MAPKAP Kinase 1b antibody [Y82] - BSA and Azide free (AB247218)

This data was developed using ab32062, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Rsk 2 / MAPKAP Kinase 1b antibody [Y82] (<a href='/en-us/products/primary-antibodies/rsk-2-mapkap-kinase-1b-antibody-y82-ab32062'>ab32062</a>) at 1/1000 dilution

All lanes:

MCF7 cell lysate

Predicted band size: 84 kDa

Observed band size: 90 kDa

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• WB

Lab

Western blot - Anti-Rsk 2 / MAPKAP Kinase 1b antibody [Y82] - BSA and Azide free (AB247218)

This data was developed using ab32062, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 cell lysate (40 μg)

Lane 2 : Rsk 2 / MAPKAP Kinase 1b knockout HAP1 cell lysate (40 μg)

Lane 3 : MCF7 cell lysate (40 μg)

Lane 4 : HeLa cell lysate (40 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab32062 observed at 88 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32062 was shown to recognize Rsk 2 / MAPKAP Kinase 1b when Rsk 2 / MAPKAP Kinase 1b knockout samples were used, along with additional cross-reactive bands. Wild-type and Rsk 2 / MAPKAP Kinase 1b knockout samples were subjected to SDS-PAGE. ab32062 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10000 dilution respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Rsk 2 / MAPKAP Kinase 1b antibody [Y82] (<a href='/en-us/products/primary-antibodies/rsk-2-mapkap-kinase-1b-antibody-y82-ab32062'>ab32062</a>)

Predicted band size: 84 kDa

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