Anti-RSK1 p90 (phospho T359) antibody [E238]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK1 p90 (phospho T359) antibody [E238] (AB32413)

ab32413, at a 1/100 dilution, staining RSK1 p90 in paraffin embedded human cervical carcinoma tissue by Immunohistochemistry.
Left image : Untreated.
Right image : Treated with Phosphatase.

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

• ELISA

Lab

ELISA - Anti-RSK1 p90 (phospho T359) antibody [E238] (AB32413)

ELISA analysis using ab32413 at a range of 0-1000 ng/ml followed by a Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L）at 1/2500 dilution. Antigen concentration : 100 ng/ml.

Antigens : Human RSK1 p90 (phospho T359 + S363) peptide, Human RSK1 p90 (phospho T359) peptide, Human RSK1 p90 (phospho S363) peptide, Human RSK1 non-phospho peptide.

• IP

Unknown

Immunoprecipitation - Anti-RSK1 p90 (phospho T359) antibody [E238] (AB32413)

Purified ab32413 at 1/100 dilution (2μg) immunoprecipitating RSK1 p90 in NIH/3T3 whole cell lysate.
Lane 1 (input) : NIH/3T3 (Mouse embryonic fibroblast) serum starved for 4h, then treated with 20% FBS for 30min whole cell lysate 10μg
Lane 2 (+) : ab32413 + NIH/3T3 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab171376 in NIH/3T3 whole cell lysate.
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
Lower bands could be cleavage product. (PMID : 29290690)

All lanes:

Immunoprecipitation - Anti-RSK1 p90 (phospho T359) antibody [E238] (ab32413)

Predicted band size: 82 kDa

false

• WB

Lab

Western blot - Anti-RSK1 p90 (phospho T359) antibody [E238] (AB32413)

Blocking/Diluting Buffer and concentration 5% NFDM/TBST

All lanes:

Western blot - Anti-RSK1 p90 (phospho T359) antibody [E238] (ab32413) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) grown in serum free media for 4 hour swhole cell lysate at 10 µg

Lane 2:

NIH/3T3 grown in serum free media for 4 hours, then treated with 20% FBS for 30minutes whole cell lysate at 10 µg

Lane 3:

NIH/3T3 grown in serum free media for 4 hours, then treated with 20% FBS for 30 minutes whole cell lysate. Then the membrane was incubated with alkaline phosphatase at 10 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

Unknown

Western blot - Anti-RSK1 p90 (phospho T359) antibody [E238] (AB32413)

All lanes:

Western blot - Anti-RSK1 p90 (phospho T359) antibody [E238] (ab32413) at 1/20000 dilution

Lane 1:

A431 cell lysate untreated.

Lane 2:

A431 cell lysate treated with EGF.

Predicted band size: 82 kDa

Observed band size: 90 kDa

false

• Dot

Unknown

Dot Blot - Anti-RSK1 p90 (phospho T359) antibody [E238] (AB32413)

Dot blot analysis of RSK1 p90 (phospho T359 + S363) phospho peptide (Lane 1), RSK1 p90 phospho T359 phospho peptide (Lane 2), RSK1 p90 phospho S363 phospho peptide (Lane 3) and RSK1 p90 non-phospho peptide (Lane 4) labeling RSK1 p90 (phospho T359 + S363) phospho peptide with purified ab32413 at a dilution of 1/1000 dilution (2.056ug/ml). A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000 dilution.

Blocking buffer : 5% NFDM/TBST. Dilution buffer : 5% NFDM /TBST .

• WB

CiteAb

Western blot - Anti-RSK1 p90 (phospho T359) antibody [E238] (AB32413)

RSK1 p90 (phospho T359) western blot using anti-RSK1 p90 (phospho T359) antibody [E238] ab32413. Publication image and figure legend from Lu, W., Jia, D., et al., 2017, Sci Rep, PubMed 28827748.

ab32413 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab32413 please see the product overview.

CSEI regulated the expressions of hematopoietic function related proteins in CHRF and K562 cells. After a 24-h CSEI (0, 0.05, 0.1 mg/ml) treatment, the protein expression levels of P-RSK1p90, ELK1 and c-Myc in CHRF cells (a) and K562 cells (b), full-length blots are presented in Supplementary Fig. S5 a, and the expression levels of GATA-1 and GATA-2 in the terminal mature CHRF cells (c) and differentiated K562 cells (d) were detected by western blot, respectively, full-length blots are presented in Supplementary Fig. S5 (b). The quantitative data of these protein levels were normalized by their GAPDH expressions and expressed as a percentage of the corresponding relative intensity of control. The data are shown as the means ± S.E.M. (n = 6). *p < 0.05, **p < 0.01 and ***p < 0.001 versus 0 mg/ml CSEI-treated cells.

false