Recombinant Anti-MCM2 (phospho S53) antibody [EP4120] (ab109133)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP4120] to MCM2 (phospho S53)
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-MCM2 (phospho S53) antibody [EP4120]
See all MCM2 primary antibodies -
Description
Rabbit monoclonal [EP4120] to MCM2 (phospho S53) -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt or IP -
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: HeLa whole cell lysate (ab150035). IHC-P:Human tonsil tissue. ICC/IF: HeLa cells.
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General notes
This product has switched from a hybridoma to recombinant production method on 9th June 2023.
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 -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP4120 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109133 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 |
---|---|---|
Flow Cyt (Intra) |
1/20.
|
|
WB | (1) |
1/1000.
|
IHC-P | (2) |
1/2400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
ICC/IF | (1) |
1/50.
|
Notes |
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Flow Cyt (Intra)
1/20. |
WB
1/1000. |
IHC-P
1/2400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/50. |
Target
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Function
Acts as component of the MCM2-7 complex (MCM complex) which is the putative replicative helicase essential for 'once per cell cycle' DNA replication initiation and elongation in eukaryotic cells. The active ATPase sites in the MCM2-7 ring are formed through the interaction surfaces of two neighboring subunits such that a critical structure of a conserved arginine finger motif is provided in trans relative to the ATP-binding site of the Walker A box of the adjacent subunit. The six ATPase active sites, however, are likely to contribute differentially to the complex helicase activity. Required for the entry in S phase and for cell division. -
Sequence similarities
Belongs to the MCM family.
Contains 1 MCM domain. -
Post-translational
modificationsPhosphorylated on Ser-108 by ATR in proliferating cells. Ser-108 proliferation is increased by genotoxic agents. Ser-40 is mediated by the CDC7-DBF4 and CDC7-DBF4B complexes, while Ser-53 phosphorylation is only mediated by the CDC7-DBF4 complex. Phosphorylation by the CDC7-DBF4 complex during G1/S phase is required for the initiation of DNA replication. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4171 Human
- Entrez Gene: 17216 Mouse
- Entrez Gene: 312538 Rat
- Omim: 116945 Human
- SwissProt: P49736 Human
- SwissProt: P97310 Mouse
- Unigene: 477481 Human
- Unigene: 16711 Mouse
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Alternative names
- BM28 antibody
- CCNL 1 antibody
- CCNL1 antibody
see all
Images
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All lanes : Anti-MCM2 (phospho S53) antibody [EP4120] (ab109133) at 1/10000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Lane 2 : HeLa whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 130 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
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All lanes : Anti-MCM2 (phospho S53) antibody [EP4120] (ab109133) at 1/10000 dilution
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg
Lane 2 : NIH/3T3 whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 130 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
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All lanes : Anti-MCM2 (phospho S53) antibody [EP4120] (ab109133) at 1/10000 dilution
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg
Lane 2 : C6 whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 130 kDa why is the actual band size different from the predicted?
Exposure time: 60 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa cells labelling MCM2 with ab109133 at 1/50 dilution (Red) compared with an isotype control (Black) and an unlabelled control (Cell without incubation with primary antibody and secondary antibody (Blue)). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) was used as the secondary antibody.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling MCM2 with ab109133 at 1/50 dilution. Confocal image showing cytoplasmic and membranous staining in subsets of HeLa cells. ab195889 AlexaFluor® 594-conjugated Anti-alpha Tubulin was used to counterstain tubulin at 1/200 dilution. The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling MCM2 with ab109133 at 1/2400 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Human tonsil tissue without alkaline phosphatase treatment (A) compared to no signal detected when treated with alkaline phosphatase (B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval Solution2) for 30 mins. Counterstained with hematoxylin.
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Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling MCM2 with ab109133 at 1/2400 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Human colon tissue without alkaline phosphatase treatment (A) compared to no signal detected when treated with alkaline phosphatase (B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval Solution2) for 30 mins. Counterstained with hematoxylin.
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Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling MCM2 with ab109133 at 1/2400 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Mouse spleen tissue without alkaline phosphatase treatment (A) compared to no signal detected when treated with alkaline phosphatase (B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval Solution2) for 30 mins. Counterstained with hematoxylin.
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Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling MCM2 with ab109133 at 1/2400 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Rat spleen tissue without alkaline phosphatase treatment (A) compared to no signal detected when treated with alkaline phosphatase (B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval Solution2) for 30 mins. Counterstained with hematoxylin.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab109133 has been referenced in 9 publications.
- Borel V et al. Disrupted control of origin activation compromises genome integrity upon destabilization of Polε and dysfunction of the TRP53-CDKN1A/P21 axis. Cell Rep 39:110871 (2022). PubMed: 35649380
- Guo Y et al. Targeting CDC7 potentiates ATR-CHK1 signaling inhibition through induction of DNA replication stress in liver cancer. Genome Med 13:166 (2021). PubMed: 34663432
- Wienert B et al. Timed inhibition of CDC7 increases CRISPR-Cas9 mediated templated repair. Nat Commun 11:2109 (2020). PubMed: 32355159
- Guarino Almeida E et al. A kinase-independent function for AURORA-A in replisome assembly during DNA replication initiation. Nucleic Acids Res 48:7844-7855 (2020). PubMed: 32652013
- Wang C et al. Inducing and exploiting vulnerabilities for the treatment of liver cancer. Nature 574:268-272 (2019). PubMed: 31578521
- Qian X et al. PTEN Suppresses Glycolysis by Dephosphorylating and Inhibiting Autophosphorylated PGK1. Mol Cell 76:516-527.e7 (2019). PubMed: 31492635
- Li X et al. Nuclear PGK1 Alleviates ADP-Dependent Inhibition of CDC7 to Promote DNA Replication. Mol Cell 72:650-660.e8 (2018). PubMed: 30392930
- Alver RC et al. Reversal of DDK-Mediated MCM Phosphorylation by Rif1-PP1 Regulates Replication Initiation and Replisome Stability Independently of ATR/Chk1. Cell Rep 18:2508-2520 (2017). PubMed: 28273463
- Garzón J et al. Shortage of dNTPs underlies altered replication dynamics and DNA breakage in the absence of the APC/C cofactor Cdh1. Oncogene 36:5808-5818 (2017). PubMed: 28604743