phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Recombinant Antibody

$25 ug total protein per lane of various lysates (see on figure) probed with phospho-ERK1 (T202\/Y204)\/ERK2 (T185\/Y187) monoclonal antibody, unconjugated (bsm-63322R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with phospho-ERK1 (T202\/Y204)\/ERK2 (T185\/Y187) monoclonal antibody, unconjugated (bsm-63322R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$The Hela (treated with 200 ng\/ml PMA for 30 minutes) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-ERK1 (T202\/Y204)\/ERK2 (T185\/Y187) antibody (bsm-63322R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

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\u2103,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 (T202\/Y204)\/ERK2 (T185\/Y187) antibody (bsm-63322R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

$The Jurkat (treated with 200 ng\/ml PMA for 30 minutes) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-ERK1 (T202\/Y204)\/ERK2 (T185\/Y187) antibody (bsm-63322R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

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\u2103,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 (T202\/Y204)\/ERK2 (T185\/Y187) antibody (bsm-63322R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

$The C6 (treated with 200 ng\/ml PMA for 30 minutes) (Rat) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-ERK1 (T202\/Y204)\/ERK2 (T185\/Y187) antibody (bsm-63322R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

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\u2103,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 (T202\/Y204)\/ERK2 (T185\/Y187) antibody (bsm-63322R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

$4% Paraformaldehyde-fixed HeLa (treated with 200 ng\/ml TPA\/PMA for 30 minutes) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-ERK1 (T202\/Y204)\/ERK2 (T185\/Y187)) monoclonal Antibody, unconjugated (bsm-63322R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.$

4% Paraformaldehyde-fixed HeLa (treated with 200 ng\/ml TPA\/PMA for 30 minutes) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-ERK1 (T202\/Y204)\/ERK2 (T185\/Y187)) monoclonal Antibody, unconjugated (bsm-63322R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C 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.

Applications

Reactivity

Human
Rat

Predicted Reactivity

Mouse
Pig

Overview
Catalog #	bsm-63322r
Product Name	phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Recombinant Antibody
Applications	WB FCM
Reactivity	Human, Rat
Predicted Reactivity	Mouse, Pig
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	A synthesized peptide derived from human ERK1 around the phosphorylation site of T202/Y204
Clonality	Recombinant
Clone #	5A12
Isotype	IgG
Concentration	1mg/ml
Purification	purified by Protein A.
Storage Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	5594
Swiss Prot	P27361
Subcellular location	Nucleus
Synonyms	MK03_HUMAN; MAPK3; MAP kinase 3; MAPK 3; ERT2; Extracellular signal-regulated kinase 1 (ERK-1); Insulin-stimulated MAP2 kinase; MAP kinase isoform p44 (p44-MAPK); Microtubule-associated protein 2 kinase; p44-ERK1; 2.7.11.24; ERK1; PRKM3; MK01_HUMAN; MAPK1; MAP kinase 1; MAPK 1; ERT1; Extracellular signal-regulated kinase 2 (ERK-2); MAP kinase isoform p42 (p42-MAPK); Mitogen-activated protein kinase 2 (MAP kinase 2 \| MAPK 2); ERK2; PRKM1; PRKM2; ERK; ERK-2; MAPK2; NS13; P42MAPK; p38; p40; p41; p41mapk; p42-MAPK; ERK-1; HS44KDAP; HUMKER1A; P44ERK1; P44MAPK; p44-MAPK; mitogen-activated protein kinase 3; ERK1 & ERK2 (phospho-T202/Y204/T183/Y185); p-ERK1 & ERK2; phospho-ERK1 & ERK2; ERK1 & ERK2 (phospho-Thr202/Tyr204/Thr183/Tyr185); p44/42
Application Dilution
WB	1:500-2000
FCM	FCM1:50-100