Recombinant Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18444] to ERK1 (phospho T202) + ERK2 (phospho T185) - BSA and Azide free
- Suitable for: IHC-P, ICC/IF, IP, WB, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free -
Description
Rabbit monoclonal [EPR18444] to ERK1 (phospho T202) + ERK2 (phospho T185) - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IF, IP, WB, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate; NIH/3T3 treated with 50 ng/ml PDGF for 40 minutes whole cell lysate; PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate. IHC-P: Human breast, placenta, breast cancer and glioma tissues; mouse kidney tissue; rat spleen tissue. ICC/IF: Jurkat cells treated with PMA treatment (200 ng/ml, 30min). IP: Jurkat treated with 200 ng/ml PMA for 30 minutes cell lysate; PC-12 treated with 200 ng/ml NGF for 4 days cell lysate.
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General notes
ab222493 is the carrier-free version of ab214036.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18444 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab222493 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 43, 41 kDa).
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Dot blot |
Use at an assay dependent concentration.
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Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 43, 41 kDa). |
Dot blot
Use at an assay dependent concentration. |
Target
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Cellular localization
ERK2: Nucleus. -
Database links
- Entrez Gene: 5594 Human
- Entrez Gene: 5595 Human
- Entrez Gene: 26413 Mouse
- Entrez Gene: 26417 Mouse
- Entrez Gene: 116590 Rat
- Entrez Gene: 50689 Rat
- Omim: 176948 Human
- Omim: 601795 Human
see all
Images
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Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Dot blot analysis of ERK1 (pT202) peptide (Lane 1), ERK1 (pT204) peptide (Lane 2), ERK1 (pT202 + pT204) peptide (Lane 3) and ERK1 non-phospho peptide (Lane 4) labelling ERK1 (pT202) with ab214036.
Exposure time: 3 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Immunohistochemical analysis of paraffin-embedded human breast tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear staining on human normal breast tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on scattered cells of human placenta is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on human breast tissue cancer is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on human glioma is observed [PMID:17487353].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on scattered cells of mouse kidney is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on scattered cells of rat spleen is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling -ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining increased after PMA treatment (200 ng/ml, 30min), and LP treatment decreased the PMA induced staining.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab214036 at 1/100 dilution followed by Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
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Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining increased after PMA treatment (200 ng/ml, 30min), and LP treatment decreased the PMA induced staining. For the “pan” antibody, the signal is unchanged after PMA treatment (200 ng/ml, 30min), and LP treatment. The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
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Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
ERK1 (phospho T202) + ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 200 ng/ml PMA for 30 minutes whole cell lysate with ab214036 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab214036 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate 10µg (Input).
Lane 2: ab214036 IP in Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214036 in Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
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Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
ERK1 (phospho T202) + ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate with ab214036 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab214036 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate 10µg (Input).
Lane 2: ab214036 IP in PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214036 in PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (2)
ab222493 has been referenced in 2 publications.
- Liu B et al. miR-181b-5p promotes proliferation and inhibits apoptosis of hypertrophic scar fibroblasts through regulating the MEK/ERK/p21 pathway. Exp Ther Med 17:1537-1544 (2019). PubMed: 30783419
- He C et al. Exopolysaccharide from Paecilomyces lilacinus modulates macrophage activities through the TLR4/NF-?B/MAPK pathway. Mol Med Rep 20:4943-4952 (2019). PubMed: 31638207