Recombinant Anti-MAPK6/ERK3 antibody [EP1720Y] - BSA and Azide free (ab247348)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1720Y] to MAPK6/ERK3 - BSA and Azide free
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-MAPK6/ERK3 antibody [EP1720Y] - BSA and Azide free
See all MAPK6/ERK3 primary antibodies -
Description
Rabbit monoclonal [EP1720Y] to MAPK6/ERK3 - BSA and Azide free -
Host species
Rabbit -
Specificity
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt or ICC/IF -
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: Wild-type HeLa, 2.4G2, HAP1, A431, C2C12, and HeLa (ab150035) whole cell lysates, and rat brain tissue lysates. IHC-P: Human kidney carcinoma and breast carcinoma tissues.
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General notes
ab247348 is the carrier-free version of ab53277.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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.
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 production
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
EP1720Y -
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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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab247348 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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WB |
Use at an assay dependent concentration. Detects a band of approximately 105 kDa (predicted molecular weight: 63 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 105 kDa (predicted molecular weight: 63 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
Phosphorylates microtubule-associated protein 2 (MAP2). May promote entry in the cell cycle. -
Tissue specificity
Highest expression in the skeletal muscle, followed by the brain. Also found in heart, placenta, lung, liver, pancreas, kidney and skin fibroblasts. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain. -
Domain
The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases. -
Post-translational
modificationsDually phosphorylated on Thr-626 and Tyr-628, which activates the enzyme. - Information by UniProt
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Database links
- Entrez Gene: 5597 Human
- Entrez Gene: 50772 Mouse
- Entrez Gene: 58840 Rat
- Omim: 602904 Human
- SwissProt: Q16659 Human
- SwissProt: Q61532 Mouse
- SwissProt: P27704 Rat
- Unigene: 411847 Human
see all -
Alternative names
- ERK-3 antibody
- ERK3 antibody
- Extracellular signal regulated kinase 3 antibody
see all
Images
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All lanes : Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MAPK6 knockout HeLa cell lysate
Lane 3 : 2.4G2 (Rat B cell lymphoma B lymphocyte) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 63 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab53277).
Lanes 1-3: Merged signal (red and green). Green - ab53277 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.
ab53277 Anti-MAPK6/ERK3 antibody [EP1720Y] was shown to specifically react with 3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264910 (knockout cell lysate ab257526) was used. Wild-type and 3 knockout samples were subjected to SDS-PAGE. ab53277 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney carcinoma tissue sections labeling MAPK6/ERK3 with Purified ab53277 at 1:100 dilution (13.52 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Purified)ImmunoHistoProbe one step HRP Polymer (ready to use) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab53277) -
All lanes : Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : MAPK6 knockout HAP1 whole cell lysate
Lane 3 : A431 whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 63 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab53277).
Lanes 1 - 4: Merged signal (red and green). Green - ab53277 observed at 90 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab53277 was shown to react with MAPK6 in HAP1 wild-type cells in Western blot. Loss of signal was observed when MAPK6 knockout sample was used. HAP1 wild-type and MAPK6 knockout whole cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab53277 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab247348 has not yet been referenced specifically in any publications.