bsm-54491R [Primary Antibody]
Phospho-Erk1(T202)+Erk2(T185) Monoclonal Antibody
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Host: Rabbit

Target Protein: Phospho-Erk1(T202)+Erk2(T185)

Modification Site: Thr202+Thr185

Clonality: Monoclonal

Isotype: IgG

Entrez Gene: 5595

Swiss Prot: P27361

Source: Synthetic phospho-Peptide corresponding to residues surrounding Thr185 of human Erk2.

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Storage: Store at -20°C for 12 months.

Background:

The protein encoded by this gene is a member of the MAPkinase family. MAP kinases, also known as extracellularsignal-regulated kinases (ERKs), act in a signaling cascade thatregulates various cellular processes such as proliferation,differentiation, and cell cycle progression in response to avariety of extracellular signals. This kinase is activated byupstream kinases, resulting in its translocation to the nucleuswhere it phosphorylates nuclear targets. Alternatively splicedtranscript variants encoding different protein isoforms have beendescribed. [provided by RefSeq, Jul 2008].

Size: 100µL

Concentration: Lot Dependent

Applications: WB(WB(1:500-2000))
FCM(FCM(1:50-100))
IHC-P(IHC-P(1:50-200))
IHC-F(IHC-F(1:50-200))
IF(IHC-P)(IF(IHC-P)(1:50-200))
IF(ICC)(IF(ICC)(1:50-200))

Predicted Molecular Weight: 42/44


Cross Reactive Species: Human
Mouse
Rat

For research use only. Not intended for diagnostic or therapeutic use.

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VALIDATION IMAGES

Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated(bsm-54491R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining


Paraformaldehyde-fixed, paraffin embedded Rat Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated(bsm-54491R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining


Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated(bsm-54491R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining


Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated(bsm-54491R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining


Paraformaldehyde-fixed, paraffin embedded Human Thyroid tumor； Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated(bsm-54491R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining


Paraformaldehyde-fixed, paraffin embedded Human Breast； Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated(bsm-54491R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining


Paraformaldehyde-fixed, paraffin embedded Mouse Placenta； Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated(bsm-54491R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining


Paraformaldehyde-fixed, paraffin embeddedRat Placenta； Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated(bsm-54491R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining


4% Paraformaldehyde-fixed Hela (treated with 200 ng/ml PMA for 30 minutes) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-ERK1 (Thr202)+ ERK2 (Thr185)) monoclonal Antibody, unconjugated (bsm-54491R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.


The Jurkat (treated with 200 ng/ml PMA for 30 minutes) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-phospho-ERK1 (Thr202)+ ERK2 (Thr185 antibody (bsm-54491R,1:100); Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488 (bs-60295G-BF488):1μg/test. Blank control (black): PBS. Acquisition of 20,000 events was performed.


The Hela (treated with 200 ng/ml PMA for 30 minutes) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-phospho-ERK1 (Thr202)+ ERK2 (Thr185 antibody (bsm-54491R,1:100); Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488 (bs-60295G-BF488):1μg/test.Blank control (black): PBS. Acquisition of 20,000 events was performed.


The C6 (treated with 200 ng/ml PMA for 30 minutes) (Rat) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-phospho-ERK1 (Thr202)+ ERK2 (Thr185 antibody (bsm-54491R,1:100); Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488 (bs-60295G-BF488):1μg/test. Blank control (black): PBS. Acquisition of 20,000 events was performed.


The NIH/3T3 (treated with 200 ng/ml PMA for 30 minutes) (M) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-phospho-ERK1 (Thr202)+ ERK2 (Thr185 antibody (bsm-54491R,1:100); Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488 (bs-60295G-BF488):1μg/test. Blank control (black): PBS. Acquisition of 20,000 events was performed.


Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated (bsm-54491R) at 1:200overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated (bsm-54491R) at 1:200overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated (bsm-54491R) at 1:200overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


The HeLa (treated with 200 ng/ml TPA/PMA for 30 minutes) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-ERK1/ERK2(T202/T185) antibody (bsm-54491R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.


Paraformaldehyde-fixed, paraffin embedded Human Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-ERK1 (Thr202)+ ERK2 (Thr185) Monoclonal Antibody, Unconjugated (bsm-54491R) at 1:200overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


25 ug total protein per lane of various lysates (see on figure) probed with phospho-ERK1/ERK2(T202/T185) monoclonal antibody, unconjugated (bsm-54491R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.


25 ug total protein per lane of various lysates (see on figure) probed with phospho-ERK1 (Thr202)+ ERK2 (Thr185) monoclonal antibody, unconjugated (bsm-54491R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.