Recombinant Anti-Mark3 antibody [EPR633Y] (ab52626)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR633Y] to Mark3
- Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Mark3 antibody [EPR633Y]
See all Mark3 primary antibodies -
Description
Rabbit monoclonal [EPR633Y] to Mark3 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, Flow Cyt (Intra), ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide within Human Mark3 aa 600-700 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa, K562 and NIH/3T3 cell lysate. ICC/IF: MCF-7 cells. Flow Cyt (intra): HeLa cells. IP: K562 cells.
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General notes
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR633Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab52626 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 |
1/1000 - 1/2000. Detects a band of approximately 86 kDa.
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IP | (1) |
1/20 - 1/60.
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Flow Cyt (Intra) |
1/30 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/50 - 1/100.
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Notes |
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WB
1/1000 - 1/2000. Detects a band of approximately 86 kDa. |
IP
1/20 - 1/60. |
Flow Cyt (Intra)
1/30 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/50 - 1/100. |
Target
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Function
Involved in the specific phosphorylation of microtubule-associated proteins for tau, MAP2 and MAP4. Phosphorylates CDC25C on 'Ser-216'. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. MARK subfamily.
Contains 1 KA1 (kinase-associated) domain.
Contains 1 protein kinase domain.
Contains 1 UBA domain. - Information by UniProt
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Database links
- Entrez Gene: 4140 Human
- Entrez Gene: 17169 Mouse
- Omim: 602678 Human
- SwissProt: P27448 Human
- SwissProt: Q03141 Mouse
- Unigene: 35828 Human
- Unigene: 260504 Mouse
- Unigene: 408955 Mouse
see all -
Alternative names
- C-TAK1 antibody
- Cdc25C associated protein kinase 1 antibody
- Cdc25C-associated protein kinase 1 antibody
see all
Images
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ab52626 (purified) at a dilution of 1/20 immunoprecipitating Mark3 in K562 whole cell lysate.
Lane 1 (input): K562 whole cell lysate (10µg)
Lane 2 (+): ab52626 + K562 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52626 in K562 whole cell lysate.
For western blotting, ab131366 VeriBlot for IP (HRP) was used for detection (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: MARK3 knockout HAP1 cell lysate (40 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: K562 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab52626 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.ab52626 was shown to recognize Mark3 when Mark3 knockout samples were used, along with additional cross-reactive bands. Wild-type and Mark3 knockout samples were subjected to SDS-PAGE. Ab52626 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1:10,000 dilution respectively and incubated overnight at 4C. 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 1:10,000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Mark3 antibody [EPR633Y] (ab52626) at 1/2000 dilution (purified)
Lane 1 : K562 whole cell lysate
Lane 2 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 86 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST -
Anti-Mark3 antibody [EPR633Y] (ab52626) at 10 µg (purified) + NIH/3T3 whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 86 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST -
Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Mark3 with purified ab52626 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
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Intracellular Flow Cytometry analysis ofHeLa cells labelling Mark3 with purified ab52626 at a dilution of 1/50 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluorr® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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Anti-Mark3 antibody [EPR633Y] (ab52626) at 1/2000 dilution (unpurified) + HeLa cell lysate at 10 µg
Secondary
HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Observed band size: 86 kDa why is the actual band size different from the predicted? -
Overlay histogram showing HeLa cells stained with unpurified ab52626 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab52626, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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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SDS download
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Datasheet download
References (2)
ab52626 has been referenced in 2 publications.
- Fredriksson K et al. Proteomic analysis of proteins surrounding occludin and claudin-4 reveals their proximity to signaling and trafficking networks. PLoS One 10:e0117074 (2015). PubMed: 25789658
- Denison FC et al. LKB1 is an essential regulator of spermatozoa release during spermiation in the mammalian testis. PLoS One 6:e28306 (2011). IP ; Mouse . PubMed: 22145035