ET1603-23

ERK2 Recombinant Rabbit Monoclonal Antibody [SZ25-01]

cat.: ET1603-23

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC, IHC-Fr
Clonality:	Monoclonal
Clone number:	SZ25-01

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 41 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human ERK2 aa 311-360 / 360.
Positive control:	HEK-293 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, C6 cell lysate, Hela, MCF-7, NIH/3T3, human pancreas tissue, mouse pancreas tissue, rat pancreas tissue, mouse hippocampus tissue, mouse cerebral cortex tissue.
Subcellular location:	Cytoplasm, Nucleus, Cytoskeleton, Membrane, Cell junction.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP IHC-Fr	1:2,000-1:5,000 1:50-1:200 1:50-1:200 1:200-1:1,000 1:1,000 Use at an assay dependent concentration. 1:100
Uniprot #:	SwissProt: P28482 Human \| P63085 Mouse \| P63086 Rat
Alternative names:	ERK 2 ERK ERK-2 ERT1 Extracellular Signal Regulated Kinase 2 Extracellular signal-regulated kinase 2 MAP kinase 1 MAP kinase 2 MAP kinase isoform p42 MAPK 1 MAPK 2 Mapk1 MAPK2 Mitogen-activated protein kinase 1 Mitogen-activated protein kinase 2 MK01_HUMAN P38 P40 P41 p42-MAPK P42MAPK PRKM1 PRKM2 protein kinase, mitogen-activated, 1 protein kinase, mitogen-activated, 2 protein tyrosine kinase ERK2

Images

Fig1: Western blot analysis of ERK2 on different lysates with Rabbit anti-ERK2 antibody (ET1603-23) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si ERK2 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 41 kDa
Observed band size: 41 kDa

Exposure time: 1 minute; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1603-23) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of ERK2 on different lysates with Rabbit anti-ERK2 antibody (ET1603-23) at 1/2,000 dilution.

Lane 1: HEK-293 cell lysate
Lane 2: HeLa cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: PC-12 cell lysate
Lane 6: C6 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 41 kDa
Observed band size: 41 kDa

Exposure time: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1603-23) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: ICC staining of ERK2 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-23, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig4: ICC staining of ERK2 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-23, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig5: ICC staining of ERK2 in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-23, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig6: Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-ERK2 antibody (ET1603-23) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1603-23) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig7: Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-ERK2 antibody (ET1603-23) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1603-23) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig8: Immunohistochemical analysis of paraffin-embedded rat pancreas tissue with Rabbit anti-ERK2 antibody (ET1603-23) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1603-23) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig9: Immunofluorescence analysis of frozen mouse hippocampus tissue labeling ERK2 with Rabbit anti-ERK2 antibody (ET1603-23). The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody ((ET1603-23, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
	Fig10: Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling ERK2 with Rabbit anti-ERK2 antibody (ET1603-23). The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody ((ET1603-23, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.

Fig11: Flow cytometric analysis of NIH/3T3 cells labeling ERK2.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1603-23, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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