Anti-MCM2 antibody (ab4461)
- Datasheet
- References (21)
- Protocols
Overview
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Product nameAnti-MCM2 antibody
See all MCM2 primary antibodies -
DescriptionRabbit polyclonal to MCM2
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Host speciesRabbit
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Tested applicationsSuitable for: IHC-Fr, ICC/IF, IHC-P, WB, IPmore details
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Species reactivityReacts with: Mouse, Human, Xenopus laevis
Predicted to work with: Chimpanzee, Gorilla, Orangutan
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Immunogen
Synthetic peptide corresponding to Human MCM2. The epitope recognized maps to a region between residues 1 and 50 of human human minichromosomal maintenance deficient 2 using the numbering given in entry NP_0045117.2 (GeneID 4171).
Database link: P49736 -
Positive control
- Breast Carcinoma, Colon Carcinoma, Small Cell Lung Cancer
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7
Preservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab4461 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 |
|---|---|---|
| IHC-Fr | Use at an assay dependent concentration. PubMed: 21068061 | |
| ICC/IF | Use a concentration of 1 µg/ml. | |
| IHC-P | Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| WB | 1/1000 - 1/10000. Predicted molecular weight: 100 kDa. | |
| IP | Use at 2-5 µg/mg of lysate. |
Target
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FunctionActs 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.
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Sequence similaritiesBelongs 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 localizationNucleus.
- Information by UniProt
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Database links
- Entrez Gene: 4171 Human
- Entrez Gene: 17216 Mouse
- Entrez Gene: 380451 Xenopus laevis
- Omim: 116945 Human
- SwissProt: P49736 Human
- SwissProt: P97310 Mouse
- SwissProt: P55861 Xenopus laevis
- Unigene: 477481 Human
see all -
Alternative names
- BM28 antibody
- CCNL 1 antibody
- CCNL1 antibody
see all
Images
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Western blot - Anti-MCM2 antibody (ab4461)All lanes : Anti-MCM2 antibody (ab4461) at 1 µg/ml
Lane 1 : NIH3T3 whole cell lysate
Lane 2 : TCMK-1 whole cell lysate
Lane 3 : 4T1 whole cell lysate
Lane 4 : CT26 whole cell lysate
Lysates/proteins at 50 µg per lane.
Predicted band size: 100 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCM2 antibody (ab4461)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human small cell lung cancer tissue labelling MCM2 with ab4461 at 1/1000 (1µg/ml). Detection: DAB. -
Immunocytochemistry/ Immunofluorescence - Anti-MCM2 antibody (ab4461)ICC/IF image of ab4461 stained MCF7 cells. The cells were 4% PFA 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 (ab4461, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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. -
Western blot - Anti-MCM2 antibody (ab4461)Sample: HeLa cell RIPA extract.
WB - 50
µ g.CoIP - 7
µ g.ab4461 incubated at 0.2
µ g/ml for 1 hour for WB.ab4461 incubated at 20
µ g for CoIP.Detection:
WB -ECL
CoIP - Coomassie Stain
Sample: HeLa cell RIPA extract.
WB - 50 µg.
CoIP - 7 µg.
ab4461 incubated at 0.2 µg/ml for 1 hour for WB.
ab4461 incubated at 20 µg for CoIP.
Detection:
WB -ECL
CoIP - Coomassie Stain -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCM2 antibody (ab4461)Ab4461 staining MCM2 in Human placenta.
Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Protocols
References
This product has been referenced in:
- Hu W et al. MiR-373-3p enhances the chemosensitivity of gemcitabine through cell cycle pathway by targeting CCND2 in pancreatic carcinoma cells. Biomed Pharmacother 105:887-898 (2018). Read more (PubMed: 30021382) »
- Hodroj D et al. An ATR-dependent function for the Ddx19 RNA helicase in nuclear R-loop metabolism. EMBO J 36:1182-1198 (2017). Read more (PubMed: 28314779) »
