ERK1/2 (3A12) Monoclonal Antibody

$Lane 1: Hela; Lane 2: SW480; Lane 3: HCT116; Lane 4: PC12 lysate probed with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:1000 overnight at 4\u00b0C followed by a conjugated secondary antibody for 60 minutes at 37\u00b0C.$

Lane 1: Hela; Lane 2: SW480; Lane 3: HCT116; Lane 4: PC12 lysate probed with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:1000 overnight at 4\u00b0C followed by a conjugated secondary antibody for 60 minutes at 37\u00b0C.

$Flow cytometric analysis of HeLa cells with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at a 1:50 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).$

Flow cytometric analysis of HeLa cells with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at a 1:50 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).

$Flow cytometric analysis of SH-SY-5Y cells with ERK1\/2 antibody at 1\/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.$

Flow cytometric analysis of SH-SY-5Y cells with ERK1\/2 antibody at 1\/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.

$IF(ICC) staining with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in HeLa cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in HeLa cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

$IF(ICC) staining with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in MCF-7 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in MCF-7 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

$IF(ICC) staining with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in NIH\/3T3 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in NIH\/3T3 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

$IF(ICC) staining with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in A549 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.$

IF(ICC) staining with ERK1\/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in A549 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100\/PBS.

$Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Pancreas lysates; Lane 3: Mouse Large intestine lysates; Lane 4: Mouse Lymph node lysates; Lane 5: Rat Cerebrum lysates; Lane 6: Rat Pancreas lysates; Lane 7: Rat Large intestine lysates; Lane 8: Rat Lymph node lysates; Lane 9: Human Hela cell lysates; Lane 10: Human SW480 cell lysates; Lane 11: Human MCF-7 cell lysates; Lane 12: Mouse NIH\/3T3 cell lysates; Lane 13: Human A549 cell lysates; Lane 14: Human A431 cell lysates probed with ERK1\/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.$

Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Pancreas lysates; Lane 3: Mouse Large intestine lysates; Lane 4: Mouse Lymph node lysates; Lane 5: Rat Cerebrum lysates; Lane 6: Rat Pancreas lysates; Lane 7: Rat Large intestine lysates; Lane 8: Rat Lymph node lysates; Lane 9: Human Hela cell lysates; Lane 10: Human SW480 cell lysates; Lane 11: Human MCF-7 cell lysates; Lane 12: Mouse NIH\/3T3 cell lysates; Lane 13: Human A549 cell lysates; Lane 14: Human A431 cell lysates probed with ERK1\/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ERK1\/2 (3A12) Monoclonal Antibody(bs-R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ERK1\/2 (3A12) Monoclonal Antibody(bs-R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Tissue\/cell:A549 cell;4% Paraformaldehyde-fixed;Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum,C-0005) at 37\u00b0C for 20 min; Antibody incubation with (ERK1\/2) monoclonal Antibody, Unconjugated (bsm-52259R) 1:100, 90 minutes at 37\u00b0C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.$

Tissue\/cell:A549 cell;4% Paraformaldehyde-fixed;Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum,C-0005) at 37\u00b0C for 20 min; Antibody incubation with (ERK1\/2) monoclonal Antibody, Unconjugated (bsm-52259R) 1:100, 90 minutes at 37\u00b0C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

$Paraformaldehyde-fixed, paraffin embedded Rat colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with ERK1\/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with ERK1\/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with ERK1\/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with ERK1\/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with ERK1\/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with ERK1\/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4\u00b0C, DAB staining.

Applications

WB
FCM
IHC-P
IF(ICC)

Reactivity

Human
Mouse
Rat
Zebrafish

Overview
Catalog #	bsm-52259R
Product Name	ERK1/2 (3A12) Monoclonal Antibody
Applications	WB, FCM, IHC-P, IF(ICC)
Reactivity	Human, Mouse, Rat, Zebrafish
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant Human ERK1 + ERK2 between 150 amino acids to C-terminus
Clonality	Monoclonal
Clone #	3A12
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	Aqueous buffered solution containing 1xTBS (pH7.4), 1%BSA, 40%Glycerol and 0.05% Sodium Azide.
Storage Condition	Store at -20°C for 12 months.
Target
Swiss Prot	P27361, P28482
Synonyms	ERK 1; ERK 2; ERK-2; ERK1; erk1/2; ERK2; ERT1; ERT2; Extracellular signalregulated kinase 1; Extracellular signal-regulated kinase 2; MAP kinase 1; MAP kinase 2; MAP kinase isoform p42; MAP kinase isoform p44; MAPK 1; MAPK 2; MAPK 3; Mapk1; MAPK2; MAPK3;Mitogen-activated protein kinase 1; Mitogenactivated protein kinase 2; MK01_HUMAN; p38; p40; p41; p42-MAPK; PRKM 2
Background	p44/42 MAP Kinase(Thr202); ERK (extracellular signal regulated kinase), also known as MAPK (mitogen activated protein kinase) has two closely related isoforms of 44 kDa and 42 kDa, respectively. These kinases belong to a family of serine/threonine kinases that are activated upon treatment of cells with a large variety of stimuli including mitogens, hormones, growth factors, cytokines, and bioactive peptides. Cell stimulation induces the activation of a signaling cascade, the downstream effects of which have been linked to the regulation of cell growth and differentiation as well as the cytoskeleton. ERK1 and ERK2 are phosphorylated within the activation loop on both a Threonine and a Tyrosine residue (within a Thr-Glu-Tyr motif) by MEKs (MAPK/ERK kinases), thereby greatly elevating the activity of ERK1&2.
Application Dilution
WB	1:300-5000
FCM	1:20-100
IHC-P	1:200-400
IF(ICC)	1:50-200

Applications

Reactivity

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