Anti-RSK1 p90 (phospho S380) antibody [E239]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RSK1 p90 (phospho S380) antibody [E239] (AB32203)

Immunocytochemistry analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 10 minutes cells labeling RSK1 p90 with Purified ab32203 at 1 : 50 dilution (2.6 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IP

Supplier Data

Immunoprecipitation - Anti-RSK1 p90 (phospho S380) antibody [E239] (AB32203)

Purified ab32203 at 1 : 20 dilution (0.8μg) immunoprecipitating RSK1 p90 in C6 whole cell lysate treated with 100nM Calyculin A for 30 minutes.

Lane 1 (input) : C6 (Rat glial tumor glial cell) whole cell lysate treated with 100nM Calyculin A for 30 minutes 10μg.
Lane 2 (+) : ab32203 + C6 whole cell lysate treated with 100nM Calyculin A for 30 minutes.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab32203 in C6 whole cell lysate treated with 100nM Calyculin A for 30 minutes.
VeriBlot for IP Detection Reagent (HRP)(ab131366) (1 : 1000 dilution) was used for Western blotting.

Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 90 kDa

All lanes:

Immunoprecipitation - Anti-RSK1 p90 (phospho S380) antibody [E239] (ab32203)

Predicted band size: 82 kDa

false

• WB

Lab

Western blot - Anti-RSK1 p90 (phospho S380) antibody [E239] (AB32203)

Blocking and dilution buffer : 5% NFDM/TBST.

Image produced using the purified version.

All lanes:

Western blot - Anti-RSK1 p90 (phospho S380) antibody [E239] (ab32203) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight whole cell lysate at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight then treated with 200nM TPA for 4 hours whole cell lysate at 15 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight then treated with 200nM TPA for 4 hours whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 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 1/20000 dilution

Predicted band size: 82 kDa

Observed band size: 90 kDa

false

• WB

Lab

Western blot - Anti-RSK1 p90 (phospho S380) antibody [E239] (AB32203)

Blocking and dilution buffer : 5% NFDM/TBST.

Image produced using the purified version.

All lanes:

Western blot - Anti-RSK1 p90 (phospho S380) antibody [E239] (ab32203) at 1/1000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast) starved for 24 hours whole cell lysate at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) starved for 24 hours then treated with 100nM Calyculin A for 30 minutes whole cell lysate at 15 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) starved for 24 hours then treated with 100nM Calyculin A for 30 minutes whole cell lysate, and then the membrane treated with Alkaline Phosphatase for 1 hour at 15 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 1/20000 dilution

Predicted band size: 82 kDa

Observed band size: 90 kDa

false

• WB

Lab

Western blot - Anti-RSK1 p90 (phospho S380) antibody [E239] (AB32203)

Blocking and dilution buffer : 5% NFDM/TBST.

Image produced using the purified version.

All lanes:

Western blot - Anti-RSK1 p90 (phospho S380) antibody [E239] (ab32203) at 1/1000 dilution

Lane 1:

Untreated PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate at 15 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma ) treated with 200ng/ml NGF for 24 hours whole cell lysate at 15 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma ) treated with 200ng/ml NGF for 24 hours whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 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 1/20000 dilution

Predicted band size: 82 kDa

Observed band size: 90 kDa

false

• Dot

Unknown

Dot Blot - Anti-RSK1 p90 (phospho S380) antibody [E239] (AB32203)

This data was produced using the unpurified format.

Dot blot analysis of RSK1 p90 (phospho S380) phospho peptide (Lane 1) and RSK1 p90 non-phospho peptide (Lane 2) labeling RSK1 p90 (phospho S380) with ab32203 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/100 000 was used as the secondary antibody.

Blocking and diluting buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.