WB
Western blot analysis of ERK1/2 on different lysates with Rabbit anti-ERK1/2 antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate, Lane 2: Jurkat cell lysate, Lane 3: HepG2 cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: PC-12 cell lysate, Lane 6: C6 cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 19 seconds; 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-ERK1/2 antibody at 1/20,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 at 1/20,000 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.IF-F
Species: Mouse, Site: Cerebral cortex, Sample: Frozen section, Antibody concentration: 1/200, Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.ICC/IF
Immunocytochemistry analysis of Jurkat cells labeling ERK1/2 with Rabbit anti-ERK1/2 antibody at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ERK1/2 antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FCM
Flow cytometric analysis of Jurkat cells labeling ERK1/2. Cells were fixed and permeabilized. Then stained with the primary antibody (red) at 1/1,000 dilution, 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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
ERK1/2 was immunoprecipitated in 0.2mg Jurkat cell lysate with Rabbit anti-ERK1/2 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-ERK1/2 antibody at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Jurkat cell lysate (input). Lane 2: Rabbit IgG instead of Rabbit anti-ERK1/2 antibody in Jurkat cell lysate. Lane 3: Rabbit anti-ERK1/2 antibody IP in Jurkat cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 43 seconds.IF-F
Species: Mouse, Site: Hippocampus, Sample: Frozen section, Antibody concentration: 1/200, Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse esophagus tissue with Rabbit anti-ERK1/2 antibody at 1/20,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 at 1/20,000 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.| Product Name | ERK1/2 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Mitogen-activated protein kinase (MAPK) signaling pathways involve two closely related MAP kinases, known as extracellular-signal-related kinase 1 (ERK 1, p44) and 2 (ERK 2, p42). Growth factors, steroid hormones, G protein-coupled receptor ligands and neurotransmitters can initiate MAPK signaling pathways. Activation of ERK 1 and ERK 2 requires phosphorylation by upstream kinases such as MAP kinasekinase (MEK), MEK kinase and Raf-1. ERK 1 and ERK 2 phosphorylation can occur at specific tyrosine and threonine sites mapping within consensus motifs that include the threonine-glutamate-tyrosine motif. ERK activation leads to dimerization with other ERKs and subsequent localization to the nucleus. Active ERK dimers phosphorylate serine and threonine residues on nuclear proteins and influence a host of responses that include proliferation, differentiation, transcription regulation and development. The human ERK 1 gene maps to chromosome 16p11.2 and encodes a 379 amino acid protein that shares 83% sequence identity to ERK 2. The human ERK2 gene maps to chromosome 22q11.21 and encodes a 360-amino acid protein. |
| Immunogen | Recombinant protein within human ERK2 aa 180-379. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIF-FIHCIPWB |
| WB:1:5000-1:20000 ICC/IF:1:2000 IHC:1:20000 IF-F:1:200-1:500 FC:1:1,000 IP:Use at an assay dependent concentration. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | ERK 1 antibody ERK 2 antibody ERK-2 antibody ERK1 antibody erk1/2 antibody ERK2 antibody ERT1 antibody ERT2 antibody Extracellular signal regulated kinase 1 antibody Extracellular signal-regulated kinase 2 antibody MAP kinase 1 antibody MAP kinase 2 antibody MAP kinase isoform p42 antibody MAP kinase isoform p44 antibody MAPK 1 antibody MAPK 2 antibody MAPK 3 antibody Mapk1 antibody MAPK2 antibody MAPK3 antibody Mitogen-activated protein kinase 1 antibody Mitogen-activated protein kinase 2 antibody MK01_HUMAN antibody p38 antibody p40 antibody p41 antibody p42-MAPK antibody PRKM 2 antibody |
|---|---|
| Molecular Weight(MW) | 42/44 kDa |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of ERK1/2 on different lysates with Rabbit anti-ERK1/2 antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate, Lane 2: Jurkat cell lysate, Lane 3: HepG2 cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: PC-12 cell lysate, Lane 6: C6 cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 19 seconds; 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-ERK1/2 antibody at 1/20,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 at 1/20,000 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.
IF-F
Species: Mouse, Site: Cerebral cortex, Sample: Frozen section, Antibody concentration: 1/200, Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.
ICC/IF
Immunocytochemistry analysis of Jurkat cells labeling ERK1/2 with Rabbit anti-ERK1/2 antibody at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ERK1/2 antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FCM
Flow cytometric analysis of Jurkat cells labeling ERK1/2. Cells were fixed and permeabilized. Then stained with the primary antibody (red) at 1/1,000 dilution, 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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
ERK1/2 was immunoprecipitated in 0.2mg Jurkat cell lysate with Rabbit anti-ERK1/2 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-ERK1/2 antibody at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Jurkat cell lysate (input). Lane 2: Rabbit IgG instead of Rabbit anti-ERK1/2 antibody in Jurkat cell lysate. Lane 3: Rabbit anti-ERK1/2 antibody IP in Jurkat cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 43 seconds.
IF-F
Species: Mouse, Site: Hippocampus, Sample: Frozen section, Antibody concentration: 1/200, Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse esophagus tissue with Rabbit anti-ERK1/2 antibody at 1/20,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 at 1/20,000 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.| Positive Control | SW480, MCF-7, Hela, PC12, HCT116, NIH/3T3, A549. |
|---|---|
| Application Notes | WB:1:5000-1:20000 ICC/IF:1:2000 IHC:1:20000 IF-F:1:200-1:500 FC:1:1,000 IP:Use at an assay dependent concentration. |
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
