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Western Blot (WB) (Figure 1. Western blot analysis of MCM2 using anti-MCM2 antibody (MBS1753967).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysatesLane 2: human CACO-2 whole cell lysatesLane 3: monkey COS-7 whole cell lysatesLane 4: human HL-60 whole cell lysatesLane 5: human HEK293 whole cell lysatesLane 6: human THP-1 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-MCM2 antigen affinity purified monoclonal antibody (Catalog # MBS1753967) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for MCM2 at approximately 125KD. The expected band size for MCM2 is at 125KD. )

Mouse MCM2 Monoclonal Antibody | anti-MCM2 antibody

Anti-MCM2 Antibody (monoclonal, 11C4)

Gene Names
MCM2; BM28; CCNL1; CDCL1; cdc19; D3S3194; MITOTIN
Reactivity
Human, Monkey
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay
Purity
Immunogen affinity purified.
Synonyms
MCM2; Monoclonal Antibody; Anti-MCM2 Antibody (monoclonal; 11C4); DNA replication licensing factor MCM2; Minichromosome maintenance protein 2 homolog; Nuclear protein BM28; BM28; CCNL1; CDCL1; KIAA0030; minichromosome maintenance complex component 2; anti-MCM2 antibody
Ordering
For Research Use Only!
Host
Mouse
Reactivity
Human, Monkey
Clonality
Monoclonal
Isotype
Mouse IgG1
Clone Number
11C4
Specificity
Mouse IgG monoclonal antibody for MCM2 detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Applicable Applications for anti-MCM2 antibody
Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM)
Application Notes
WB: 0.1-0.5ug/ml|Human, Monkey|
IHC-P: 0.5-1ug/ml|Human|
ICC/IF: 4ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Immunogen
E Coli-derived human MCM2 recombinant protein (Position: S393-R850).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of MCM2 using anti-MCM2 antibody (MBS1753967).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysatesLane 2: human CACO-2 whole cell lysatesLane 3: monkey COS-7 whole cell lysatesLane 4: human HL-60 whole cell lysatesLane 5: human HEK293 whole cell lysatesLane 6: human THP-1 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-MCM2 antigen affinity purified monoclonal antibody (Catalog # MBS1753967) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for MCM2 at approximately 125KD. The expected band size for MCM2 is at 125KD. )

Western Blot (WB) (Figure 1. Western blot analysis of MCM2 using anti-MCM2 antibody (MBS1753967).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysatesLane 2: human CACO-2 whole cell lysatesLane 3: monkey COS-7 whole cell lysatesLane 4: human HL-60 whole cell lysatesLane 5: human HEK293 whole cell lysatesLane 6: human THP-1 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-MCM2 antigen affinity purified monoclonal antibody (Catalog # MBS1753967) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for MCM2 at approximately 125KD. The expected band size for MCM2 is at 125KD. )

Immunofluorescence (IF)

(Figure 2. IF analysis of MCM2 using anti-MCM2 antibody (MBS1753967).MCM2 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 4μg/mL mouse anti-MCM2 Antibody (MBS1753967) overnight at 4 degree C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using DyLight®488 Conjugated Avidin (BA1128). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Immunofluorescence (IF) (Figure 2. IF analysis of MCM2 using anti-MCM2 antibody (MBS1753967).MCM2 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 4μg/mL mouse anti-MCM2 Antibody (MBS1753967) overnight at 4 degree C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using DyLight®488 Conjugated Avidin (BA1128). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Immunohistochemistry (IHC)

(Figure 3. IHC analysis of MCM2 using anti-MCM2 antibody (MBS1753967).MCM2 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-MCM2 Antibody (MBS1753967) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )

Immunohistochemistry (IHC) (Figure 3. IHC analysis of MCM2 using anti-MCM2 antibody (MBS1753967).MCM2 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-MCM2 Antibody (MBS1753967) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )

Immunohistochemistry (IHC)

(Figure 4. IHC analysis of MCM2 using anti-MCM2 antibody (MBS1753967).MCM2 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-MCM2 Antibody (MBS1753967) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )

Immunohistochemistry (IHC) (Figure 4. IHC analysis of MCM2 using anti-MCM2 antibody (MBS1753967).MCM2 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-MCM2 Antibody (MBS1753967) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )

Immunofluorescence (IF)

(Figure 5. IF analysis of MCM2 using anti- MCM2 antibody (MBS1753967).MCM2 was detected in immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4μg/mL mouse anti- MCM2 Antibody (MBS1753967) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Immunofluorescence (IF) (Figure 5. IF analysis of MCM2 using anti- MCM2 antibody (MBS1753967).MCM2 was detected in immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4μg/mL mouse anti- MCM2 Antibody (MBS1753967) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Flow Cytometry (FC/FACS)

(Figure 6. Flow Cytometry analysis of HL-60 cells using anti- MCM2 antibody (MBS1753967).Overlay histogram showing HL-60 cells stained with MBS1753967 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-MCM2 Antibody (MBS1753967, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 6. Flow Cytometry analysis of HL-60 cells using anti- MCM2 antibody (MBS1753967).Overlay histogram showing HL-60 cells stained with MBS1753967 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-MCM2 Antibody (MBS1753967, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-MCM2 antibody
MCM2(MINICHROMOSOME MAINTENANCE, S. CEREVISIAE, HOMOLOG OF, 2), also known as MITOTIN, CDCL1 or BM28, is a human nuclear protein that plays an important role in 2 crucial steps of the cell cycle, namely, onset of DNA replication and cell division. And it is similar to members of the family of early S-phase proteins. The MCM2 gene is mapped to 3q21. 3. The hexameric protein complex formed by MCM proteins is a key component of the pre-replication complex (pre-RC) and may be involved in the formation of replication forks and in the recruitment of other DNA replication related proteins. In the G0 stage, the MCM2 and MCM5 proteins were much less abundant than the MCM7 and MCM3 proteins, which suggests that the MCM proteins are not present in stoichiometric amounts and that only a proportion of these molecules actively participate in cell cycle regulation as part of MCM complexes.
References
1. Blow, J. J., Laskey, R. A. A role for the nuclear envelope in controlling DNA replication within the cell cycle. Nature 332: 546-548, 1988.
2. Burkhart, R., Schulte, D., Hu, D., Musahl, C., Gohring, F., Knippers, R. Interactions of human nuclear proteins P1Mcm3 and P1Cdc46. Europ. J. Biochem. 228: 431-438, 1995.
3. Todorov IT, Lavigne J, Sakr F, Kaneva R, Foisy S, Bibor-Hardy V (Jul 1991). "Nuclear matrix protein mitotin messenger RNA is expressed at constant levels during the cell cycle". Biochem Biophys Res Commun 177 (1): 395-400.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
101,896 Da
NCBI Official Full Name
DNA replication licensing factor MCM2
NCBI Official Synonym Full Names
minichromosome maintenance complex component 2
NCBI Official Symbol
MCM2
NCBI Official Synonym Symbols
BM28; CCNL1; CDCL1; cdc19; D3S3194; MITOTIN
NCBI Protein Information
DNA replication licensing factor MCM2; cyclin-like 1; nuclear protein BM28; cell devision cycle-like 1; minichromosome maintenance protein 2 homolog; minichromosome maintenance deficient 2 (mitotin); MCM2 minichromosome maintenance deficient 2, mitotin
UniProt Protein Name
DNA replication licensing factor MCM2
UniProt Gene Name
MCM2
UniProt Synonym Gene Names
BM28; CCNL1; CDCL1; KIAA0030
UniProt Entry Name
MCM2_HUMAN

NCBI Description

The protein encoded by this gene is one of the highly conserved mini-chromosome maintenance proteins (MCM) that are involved in the initiation of eukaryotic genome replication. The hexameric protein complex formed by MCM proteins is a key component of the pre-replication complex (pre_RC) and may be involved in the formation of replication forks and in the recruitment of other DNA replication related proteins. This protein forms a complex with MCM4, 6, and 7, and has been shown to regulate the helicase activity of the complex. This protein is phosphorylated, and thus regulated by, protein kinases CDC2 and CDC7. Multiple alternatively spliced transcript variants have been found, but the full-length nature of some variants has not been defined. [provided by RefSeq, Oct 2012]

Uniprot Description

MCM2: a mini-chromosome maintenance protein, essential for the initiation of eukaryotic genome replication. Allows DNA to undergo a single round of replication per cell cycle. Required for the entry in S phase and for cell division.

Protein type: DNA-binding; EC 3.6.4.12

Chromosomal Location of Human Ortholog: 3q21

Cellular Component: microtubule cytoskeleton; nucleoplasm; MCM complex; cytoplasm; chromatin; nuclear origin of replication recognition complex; nucleus

Molecular Function: protein binding; DNA helicase activity; DNA binding; histone binding; metal ion binding; DNA replication origin binding; ATP binding

Biological Process: DNA unwinding during replication; nucleosome assembly; DNA replication initiation; DNA strand elongation during DNA replication; mitotic cell cycle; cell cycle; DNA replication; G1/S transition of mitotic cell cycle

Research Articles on MCM2

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Product Notes

The MCM2 mcm2 (Catalog #AAA1753967) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-MCM2 Antibody (monoclonal, 11C4) reacts with Human, Monkey and may cross-react with other species as described in the data sheet. AAA Biotech's MCM2 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM). WB: 0.1-0.5ug/ml|Human, Monkey| IHC-P: 0.5-1ug/ml|Human| ICC/IF: 4ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the MCM2 mcm2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "MCM2, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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