Anti-ERK1 (phospho T202) + ERK2 (phospho T185) 抗体 [EPR19401]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling ERK2 (phospho T185) with ab201015 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human breast is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling ERK1 (phospho T202) and ERK2 (phospho T185)

ERK1 (phospho T202) + ERK2 (phospho T185) with ab201015 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing staining on M phase cells (PMID : 26529125). After PMA treatment (200 ng/ml, 30min), the staining was increased, and LP treatment decreased the PMA induced staining.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab201015 at 1/500 dilution followed by ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling ERK2 (phospho T185) with ab201015 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear with weak cytoplasm staining on Human glioma is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50ng/ml PDGF for 40min whole cell lysate with ab201015 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201015 at 1/1000 dilution. VeriBlot for IP Detection Reaction (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate, 10μg (Input).

Lane 2 : ab201015 IP in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab201015 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

false

• IP

Supplier Data

Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 100ng/ml NGF for 10min whole cell lysate with ab201015 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201015 at 1/1000 dilution. VeriBlot for IP Detection Reaction (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : PC-12 treated with 100ng/ml NGF for 10min whole cell lysate, 10μg (Input).

Lane 2 : ab201015 IP in PC-12 treated with 100ng/ml NGF for 10min whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab201015 in PC-12 treated with 100ng/ml NGF for 10min whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

false

• WB

Supplier Data

Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) at 1/1000 dilution

Lane 1:

Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 2:

Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 200 ng/ml PMA for 30 minutes whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 42 kDa,44 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

Blocking/Dilution buffer : 5% NFDM/TBST.

The induction conditions refer to PMID : 12454035; PMID : 17026715; PMID : 22206868.

All lanes:

Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50 ng/ml PDGF for 40 minutes whole cell lysate at 20 µg

Lane 3:

Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

Lane 4:

PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 42 kDa,44 kDa

false

Exposure time: 10s

• Dot

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Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

Dot blot analysis of ERK2 (phospho T185) labeled with ab201015 at 1/1000 dilution. **Lane 1 : ** ERK2 (pT185) phospho peptide : DHTGFLT(p)EYVATR aa179-191 peptide. **Lane 2 : ** ERK2 Non-phospho peptide : DHTGFLTEYVATR aa179-191 peptide. **Lane 3 : ** ERK2 (pY187) phospho peptide : DHTGFLTEY(p)VATR aa179-191 peptide. Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051) at 1/100000 dilution was used as secondary antibody. **Blocking/Dilution buffer : ** 5% NFDM/TBST. **Exposure time : ** 3 minutes.

• WB

CiteAb

Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (AB201015)

Western Blotting using Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401], ab201015. Publication image from He, J. et al., 2020, Mol Cancer, 31992303. Legend direct from paper.

Inhibition of phosphorylation of p38/MAPK reverses cell survival induced by circ-MAPK4. (a) U373 cells transfected with siRNA negative control, circ-MAPK4 siRNA-1 and siRNA-2 were treated with or without p-p38/MAPK inhibitor (SB203580). Inhibition efficiency of p-p38/MAPK inhibitor was accessed by testing phosphorylation and total protein levels of p38/MAPK and ERK using western blot assays. (b, c) CCK-8 and colony formation assays were performed to reveal cell survival of these U373 cells. (d) Apoptosis assays were performed to reveal apoptosis levels of these U373 cells. (e) The western blot assays were used to evaluate effect of p-p38/MAPK inhibitor on protein expression levels of cleaved form of caspase-9, caspase-7, caspase-3, PARP1 in these U373 cells. The data were summarized as bar graph. Data are the means ± SEM of three experiments. *P < 0.05; **P < 0.01; ***P < 0.001

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