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Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) is a rabbit monoclonal antibody that is used to detect ERK1 + ERK2 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, ICC/IF. Suitable for Human, Mouse, Rat samples.



- Specificity confirmed with ERK1 + ERK2 knockout cell line validation


Images

Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (AB184699), expandable thumbnail
  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (AB184699), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (AB184699), expandable thumbnail
  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (AB184699), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (AB184699), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)
Human
Expected
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Rat
Tested
Tested
Expected
Expected
Recombinant full length protein - Human
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species
Rat
Dilution info
1/70
Notes

-

Expected
Expected

Species
Mouse, Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Recombinant full length protein - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Recombinant full length protein - Human
Dilution info
1/10000
Notes

-

Species
Mouse
Dilution info
1/10000
Notes

-

Species
Rat
Dilution info
1/10000
Notes

-

Species
Human
Dilution info
1/10000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/250
Notes

-

Expected
Expected

Species
Rat, Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Recombinant full length protein - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/440
Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species
Rat, Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Recombinant full length protein - Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

6 products for Alternative Product

2 products for Alternative Version

Target data

Function

Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. MAPK1/ERK2 and MAPK3/ERK1 are the 2 MAPKs which play an important role in the MAPK/ERK cascade. They participate also in a signaling cascade initiated by activated KIT and KITLG/SCF. Depending on the cellular context, the MAPK/ERK cascade mediates diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. The MAPK/ERK cascade also plays a role in initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors. About 160 substrates have already been discovered for ERKs. Many of these substrates are localized in the nucleus, and seem to participate in the regulation of transcription upon stimulation. However, other substrates are found in the cytosol as well as in other cellular organelles, and those are responsible for processes such as translation, mitosis and apoptosis. Moreover, the MAPK/ERK cascade is also involved in the regulation of the endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC); as well as in the fragmentation of the Golgi apparatus during mitosis. The substrates include transcription factors (such as ATF2, BCL6, ELK1, ERF, FOS, HSF4 or SPZ1), cytoskeletal elements (such as CANX, CTTN, GJA1, MAP2, MAPT, PXN, SORBS3 or STMN1), regulators of apoptosis (such as BAD, BTG2, CASP9, DAPK1, IER3, MCL1 or PPARG), regulators of translation (such as EIF4EBP1 and FXR1) and a variety of other signaling-related molecules (like ARHGEF2, DCC, FRS2 or GRB10). Protein kinases (such as RAF1, RPS6KA1/RSK1, RPS6KA3/RSK2, RPS6KA2/RSK3, RPS6KA6/RSK4, SYK, MKNK1/MNK1, MKNK2/MNK2, RPS6KA5/MSK1, RPS6KA4/MSK2, MAPKAPK3 or MAPKAPK5) and phosphatases (such as DUSP1, DUSP4, DUSP6 or DUSP16) are other substrates which enable the propagation the MAPK/ERK signal to additional cytosolic and nuclear targets, thereby extending the specificity of the cascade. Mediates phosphorylation of TPR in response to EGF stimulation. May play a role in the spindle assembly checkpoint. Phosphorylates PML and promotes its interaction with PIN1, leading to PML degradation. Phosphorylates CDK2AP2 (By similarity). Acts as a transcriptional repressor. Binds to a [GC]AAA[GC] consensus sequence. Repress the expression of interferon gamma-induced genes. Seems to bind to the promoter of CCL5, DMP1, IFIH1, IFITM1, IRF7, IRF9, LAMP3, OAS1, OAS2, OAS3 and STAT1. Transcriptional activity is independent of kinase activity.

Additional Targets

MAPK3

Alternative names

ERK2, PRKM1, PRKM2, MAPK1, Mitogen-activated protein kinase 1, MAP kinase 1, MAPK 1, ERT1, Extracellular signal-regulated kinase 2, MAP kinase isoform p42, Mitogen-activated protein kinase 2, ERK-2, p42-MAPK, MAP kinase 2, MAPK 2

Recommended products

Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) is a rabbit monoclonal antibody that is used to detect ERK1 + ERK2 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, ICC/IF. Suitable for Human, Mouse, Rat samples.



- Specificity confirmed with ERK1 + ERK2 knockout cell line validation

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR17526
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

What is this antibody validated in?


Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunocytochemistry/immunofluorescence (ICC/IF), in Human, Mouse, Rat samples.

What is the molecular weight of ERK1 + ERK2?


Anti-ERK1 + ERK2 [EPR17526] (ab184699) specifically detects a band for ERK1 + ERK2 (UniProt: P28482) at a molecular weight of 43, 41kDa.

Recommended positive controls


WB: Human ERK1 full length recombinant protein; Human ERK2 full length recombinant protein; A431, Jurkat, HeLa, HepG2, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal brain, fetal heart and fetal kidney lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart, kidney and spleen lysates.ICC/IF: HeLa cells.Flow Cyt (intra): A431 cells.IP. PC-12 whole cell extract.

Trusted by the scientific community


Anti-ERK1 + ERK2 [EPR17526] (ab184699) was first used in a scientific publication in 2015 and has been cited over 250 times in peer-reviewed journals.

Trial sizes available!


Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies. Specificity confirmed


The specificity of Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) has been confirmed by Western blot testing in MAPK1 Knockout HeLa cell line, Human MAPK1 (ERK2) knockout HeLa cell line ab265052.



Other related products


We have a range of other formats of antibody clone [EPR17526] also available for your convenience:
ab184699, Carrier free - Anti-ERK1 antibody [EPR17526] - BSA and Azide free (Detector) ab242993



Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

ERK1 and ERK2 also known as MAPK3 and MAPK1 respectively are protein kinases involved in the mitogen-activated protein kinase (MAPK) signaling pathway. They share high sequence identity and exhibit similar functions. ERK1 has a molecular weight of approximately 44 kDa while ERK2 weighs around 42 kDa. Both are expressed ubiquitously in various tissues playing roles in diverse cellular processes. These proteins often detected through ERK1/2 western blot analyses present similar ERK protein size and ERK molecular weight characteristics relevant during protein expression studies.

Biological function summary

ERK1 and ERK2 play key roles in cell proliferation differentiation and survival. They form part of a cascade that includes upstream activators such as MEK1/2 and downstream targets including transcription factors. As components of the MAPK signaling complex ERK1/2 regulate gene expression through phosphorylation events impacting cellular responses to various stimuli. Their activation often hinges on growth factors cytokines and stress signals facilitating cellular adaptation to environmental changes.

Pathways

Regarding pathways ERK1/2 sit within the MAPK/ERK pathway and are significant in the Ras/Raf/MEK/ERK cascade one of the foremost signaling mechanisms in cells. They interact with several proteins including Ras and Raf which modulate their activation. This pathway is important for transmitting signals from the cell surface to the DNA in the cell nucleus impacting gene regulation and cell fate decisions. ERK1/2 proteins therefore serve as critical nodes linking extracellular signals to cellular responses ensuring balanced cell function.

Associated diseases and disorders

ERK1/2 play significant roles in cancer and neurodegenerative diseases. Their overactivation is often linked to oncogenic processes contributing to cell proliferation in cancers. Mutations or dysregulation within the MAPK/ERK pathway including ERK1/2 frequently associate with tumorigenesis. Additionally in neurodegenerative disorders like Alzheimer's disease altered ERK1/2 activity may impact neuronal survival and function often through interaction with amyloid-beta and tau proteins further illustrating their importance in disease pathophysiology.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    44kDa band represents ERK1. 42kDa band represents ERK2.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    Lane 1: Mouse brain lysates at 10 µg

    Lane 2: Mouse heart lysates at 10 µg

    Lane 3: Mouse kidney lysates at 10 µg

    Lane 4: Mouse spleen lysates at 10 µg

    Lane 5: Rat brain lysates at 10 µg

    Lane 6: Rat heart lysates at 10 µg

    Lane 7: Rat kidney lysates at 10 µg

    Lane 8: Rat spleen lysates at 10 µg

    Lane 9: C6 (Rat glial tumor cells) whole cell lysates at 10 µg

    Lane 10: RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates at 10 µg

    Lane 11: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 10 µg

    Lane 12: NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 41 kDa

    Observed band size: 42 kDa, 44 kDa

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Lanes 1-2: Merged signal (red and green). Green - ab184699 observed at 44 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.

    ab184699 Anti-ERK1 + ERK2 antibody [EPR17526] was shown to specifically react with ERK2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MAPK1 (ERK2) knockout HeLa cell line ab265052 (knockout cell lysate Human MAPK1 (ERK2) knockout HeLa cell lysate ab257525) was used. Wild-type and ERK2 knockout samples were subjected to SDS-PAGE. ab184699 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 10000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: MAPK1 knockout HeLa cell lysate at 20 µg

    Lane 2: Western blot - Human MAPK1 (ERK2) knockout HeLa cell line (Human MAPK1 (ERK2) knockout HeLa cell line ab265052)

    Performed under reducing conditions.

    Predicted band size: 41 kDa

    Observed band size: 44 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail
    This image is courtesy of a customer review submitted by Kirk Mcmanus.

    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    ab184699 staining ERK1 + ERK2 in HeLa cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. A Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used as the secondary antibody.

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Recombinant full length ERK1 protein (ab43623) contains aa1-379.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    All lanes: Recombinant Human ERK1 protein (ab43623) (ab43623) at 0.01 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 41 kDa

    Observed band size: 44 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ERK1 + ERK2 with ab184699 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing both nuclear and cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab184699 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Recombinant full length ERK2 protein (Recombinant Human ERK2 protein ab43625) contains aa1-360.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    All lanes: Western blot - Recombinant Human ERK2 protein (Recombinant Human ERK2 protein ab43625) at 0.01 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 41 kDa, 52 kDa

    Observed band size: 42 kDa

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    44kDa band represents ERK1. 42kDa band represents ERK2.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/50000 dilution

    Lane 1: A431 (Human epidermoid carcinoma) whole cell lysates at 20 µg

    Lane 2: Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates at 20 µg

    Lane 3: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 20 µg

    Lane 4: HepG2 (Human liver hepatocellular carcinoma) whole cell lysates at 20 µg

    Lane 5: Human fetal brain lysates at 20 µg

    Lane 6: Human fetal heart lysates at 20 µg

    Lane 7: Human fetal kidney lysates at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 41 kDa

    Observed band size: 42 kDa, 44 kDa

  • Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    ERK1 + ERK2 were immunoprecipitated from 1mg of PC-12 (Rat adrenal gland pheochromocytoma) whole cell extract with ab184699 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab184699 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: PC-12 whole cell extract. Lane 2: PBS instead of PC-12 whole cell extract.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    44kDa band represents ERK1. 42kDa band represents ERK2.

    All lanes: Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Predicted band size: 41 kDa

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    44kDa band represents ERK1. 42kDa band represents ERK2.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    Lane 1: Human fetal brain lysates at 10 µg

    Lane 2: Human fetal heart lysates at 10 µg

    Secondary

    All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 41 kDa

    Observed band size: 42 kDa, 44 kDa

  • Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Intracellular flow cytometric analysis of A431 (Human epidermoid carcinoma) cells labeling ERK1 + ERK2 with ab184699 at 1/440 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 staining ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) in untreated, PMA treated and PMA + LP treated HeLa cells. The cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 at 5µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

    The image shows increased nuclear staining after 24hr serum starvation followed by treatment with PMA (200nM, 15min) of HeLa cells. The Lambda Phoshatase treatment then removes all staining of antibody with no phospho ERK1/2 remaining.

    ab184699 was used as a Pan ERK1 + ERK2 control for Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063. The results showed nuclear staining on untreated, with no increase after PMA treatment and no reduction after PMA + LP treated in HeLa cells.

  • Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 staining ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) in untreated, PMA treated and PMA + LP treated NIH/3T3 cells. The cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 at 5µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

    Also suitable in cells fixed with 4% paraformaldehyde (10 min).

    The image shows increased nuclear staining after 24hr serum starvation followed by treatment with PMA (200nM, 15min) of NIH/3T3 cells. The Lambda Phoshatase treatment then removes all staining of antibody with no phospho ERK1/2 remaining.

    ab184699 was used as a Pan ERK1 + ERK2 control for Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063. The results showed nuclear staining on untreated, with no increase after PMA treatment and no reduction after PMA + LP treated in NIH/3T3 cells.

  • Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699), expandable thumbnail

    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699)

    ERK1 + ERK2 Western blot staining using rabbit Anti-ERK1 + ERK2 antibody

    Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 was shown to specifically react with ERK1/2 (pT202+ Y204). The signal was significantly reduced after treat with 10 uM U0126.

    Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% BSA in TBS-0.1 % Tween® 20 (TBS-T).

    Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063, ab184699 and Anti-Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4°C at 1/1000, 1/10000 and 1/5000 dilutions, respectively.

    Blots were developed with Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] (Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063) at 1/1000 dilution

    Lane 1: HeLa cells, serum-starved overnight, treated with U0126 (10 uM, 60 min) at 20 µg

    Lane 2: HeLa cells, serum-starved overnight, treated with TPA (200 nM, 15 min) at 20 µg

    Lane 3: NIH3T3 cells, serum-starved overnight, treated with U0126 (10 uM, 60 min) at 20 µg

    Lane 4: NIH3T3 cells, serum-starved overnight, treated with TPA (200 nM, 15 min) at 20 µg

    Lane 5: A10 cells, serum-starved overnight, treated with U0126 (10 uM, 60 min) at 20 µg

    Lane 6: A10 cells, serum-starved overnight, treated with TPA (200 nM, 15 min) at 20 µg

    Secondary

    Lanes 1 - 6: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

    Lanes 1 - 6: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 42 kDa, 44 kDa, 50 kDa

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