Recombinant Anti-CMT2 antibody [EPR9584] (ab150363)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR9584] to CMT2
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CMT2 antibody [EPR9584]
See all CMT2 primary antibodies -
Description
Rabbit monoclonal [EPR9584] to CMT2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide within Human CMT2 aa 250 to the C-terminus. The exact sequence is proprietary.
Database link: Q15013 -
Positive control
- Recombinant Human CMT2 (ab123183) can be used as a positive control in WB. HepG2, A431, and HeLa cell lysates, Human brain and Human cervical carcinoma tissues This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HeLa. WB: Wild-type HeLa cell lysate. THP-1, HepG2 and MDA-MB-231 cell lysate.
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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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR9584 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab150363 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) |
1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 31 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF |
1/100 - 1/250.
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Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 31 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
Target
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Function
May function to silence the spindle checkpoint and allow mitosis to proceed through anaphase by binding MAD2L1 after it has become dissociated from the MAD2L1-CDC20 complex. -
Sequence similarities
Belongs to the MAD2L1BP family. -
Developmental stage
During the cell cycle, levels increase and then remain constant until late mitosis after which they drop. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Nucleus. Cytoplasm > cytoskeleton > spindle. During early mitosis, unevenly distributed throughout the nucleoplasm. From metaphase to anaphase, concentrated on the spindle. - Information by UniProt
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Database links
- Entrez Gene: 9587 Human
- Entrez Gene: 66591 Mouse
- Entrez Gene: 316237 Rat
- SwissProt: Q15013 Human
- SwissProt: Q9DCX1 Mouse
- Unigene: 122346 Human
- Unigene: 715485 Human
- Unigene: 272024 Mouse
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Alternative names
- Caught by MAD2 protein antibody
- CMT2 antibody
- dJ261G23.1 antibody
see all
Images
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All lanes : Anti-CMT2 antibody [EPR9584] (ab150363) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : Human MAD2L1BP (CMT2) knockout HeLa cell lysate (ab257510)
Lane 3 : THP-1 cell lysate
Lane 4 : HepG2 cell lysate
Lane 5 : MDA-MB-231 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CMT2 antibody [EPR9584] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab150363 was shown to bind specifically to CMT2. A band was observed at 35 kDa in wild-type HeLa cell lysates with no signal observed at this size in MAD2L1BP knockout cell line ab265854 (knockout cell lysate ab257510). To generate this image, wild-type and MAD2L1BP knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-CMT2 antibody [EPR9584] (ab150363) at 1/1000 dilution
Lane 1 : HepG2 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 31 kDa
Observed band size: 34 kDa why is the actual band size different from the predicted? -
ab150363 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab150363 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Immunohistochemical analysis of paraffin embedded Human brain tissue labelling CMT2 binding protein with ab150363 antibody at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin embedded Human cervical carcinoma tissue labelling CMT2 binding protein with ab150363 antibody at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab150363 has been referenced in 4 publications.
- Richeson KV et al. Paradoxical mitotic exit induced by a small molecule inhibitor of APC/CCdc20. Nat Chem Biol 16:546-555 (2020). PubMed: 32152539
- Shin HJ et al. p31comet-Induced Cell Death Is Mediated by Binding and Inactivation of Mad2. PLoS One 10:e0141523 (2015). WB . PubMed: 26544187
- de Lange J et al. Defective sister chromatid cohesion is synthetically lethal with impaired APC/C function. Nat Commun 6:8399 (2015). WB . PubMed: 26423134
- Date DA et al. Phosphorylation regulates the p31Comet-mitotic arrest-deficient 2 (Mad2) interaction to promote spindle assembly checkpoint (SAC) activity. J Biol Chem 289:11367-73 (2014). PubMed: 24596092