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Anti-MCM2 Antibody Picoband™ (monoclonal, 11C4)

Figure 1. Western blot analysis of MCM2 using anti-MCM2 antibody (M00374-1).
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 lysates,
Lane 2: human CACO-2 whole cell lysates,
Lane 3: monkey COS-7 whole cell lysates,
Lane 4: human HL-60 whole cell lysates,
Lane 5: human HEK293 whole cell lysates,
Lane 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 # M00374-1) at 0.5 µg/mL overnight at 4°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 # EK1001) with Tanon 5200 system. A specific band was detected for MCM2 at approximately 125KD. The expected band size for MCM2 is at 125KD.
Figure 2. IF analysis of MCM2 using anti-MCM2 antibody (M00374-1).
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 4 µg/mL mouse anti-MCM2 Antibody (M00374-1) overnight at 4°C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37°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.
Figure 3. IHC analysis of MCM2 using anti-MCM2 antibody (M00374-1).
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 1 µg/ml mouse anti-MCM2 Antibody (M00374-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 4. IHC analysis of MCM2 using anti-MCM2 antibody (M00374-1).
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 1 µg/ml mouse anti-MCM2 Antibody (M00374-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Figure 5. IF analysis of MCM2 using anti-MCM2 antibody (M00374-1).
MCM2 was detected in immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4 µg/mL mouse anti-MCM2 Antibody (M00374-1) overnight at 4°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°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 6. Flow Cytometry analysis of HL-60 cells using anti-MCM2 antibody (M00374-1).
Overlay histogram showing HL-60 cells stained with M00374-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-MCM2 Antibody (M00374-1, 1 µg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC, FC
Primary Accession	P49736
Host	Mouse
Isotype	Mouse IgG1
Reactivity	Human
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-MCM2 Antibody Picoband™ (monoclonal, 11C4) . Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human.
Reconstitution	Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Additional Information
Gene ID	4171
Other Names	DNA replication licensing factor MCM2, 3.6.4.12, Minichromosome maintenance protein 2 homolog, Nuclear protein BM28, MCM2 (HGNC:6944)
Calculated MW	101896 Da
Application Details	Western blot, 0.1-0.5 µg/ml, Human, Monkey Immunohistochemistry (Paraffin-embedded Section), 0.5-1 µg/ml, Human Immunocytochemistry/Immunofluorescence, 4 µg/ml, Human Flow Cytometry, 1-3 µg/1x10^6 cells, Human
Source	Eukaryota
Contents	Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clone Names	Clone: 11C4
Immunogen	E.coli-derived human MCM2 recombinant protein (Position: S393-R850).
Purification	Immunogen affinity purified.
Storage	Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	MCM2 (HGNC:6944)
Function	Acts as a component of the MCM2-7 complex (MCM complex) which is the replicative helicase essential for 'once per cell cycle' DNA replication initiation and elongation in eukaryotic cells. Core component of CDC45-MCM-GINS (CMG) helicase, the molecular machine that unwinds template DNA during replication, and around which the replisome is built (PubMed:32453425, PubMed:34694004, PubMed:34700328, PubMed:35585232). 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 (PubMed:32453425). Required for the entry in S phase and for cell division (PubMed:8175912). Plays a role in terminally differentiated hair cells development of the cochlea and induces cells apoptosis (PubMed:26196677).
Cellular Location	Nucleus. Chromosome. Note=Associated with chromatin before the formation of nuclei and detaches from it as DNA replication progresses. {ECO:0000250\|UniProtKB:P55861}

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

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.

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¥ 3,367.00

Cat# ABO14965

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100 µg

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Anti-MCM2 Antibody Picoband™ (monoclonal, 11C4)

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