Western Blot (WB)
(Figure 1. Western blot analysis of MCM5 using anti-MCM5 antibody (MBS1754026).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 MCF-7 whole cell lysatesLane 2: rat thymus tissue lysatesLane 3: rat RH-35 whole cell lysatesLane 4: mouse thymus tissue 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- MCM5 antigen affinity purified monoclonal antibody (Catalog # MBS1754026) 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 MCM5 at approximately 97KD. The expected band size for MCM5 is at 97KD. )
Mouse MCM5 Monoclonal Antibody | anti-MCM5 antibody
Anti-MCM5 Antibody (monoclonal, 4G10)
IHC-P: 2-5ug/ml|Human|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of MCM5 using anti-MCM5 antibody (MBS1754026).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 MCF-7 whole cell lysatesLane 2: rat thymus tissue lysatesLane 3: rat RH-35 whole cell lysatesLane 4: mouse thymus tissue 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- MCM5 antigen affinity purified monoclonal antibody (Catalog # MBS1754026) 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 MCM5 at approximately 97KD. The expected band size for MCM5 is at 97KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of MCM5 using anti-MCM5 antibody (MBS1754026).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 MCF-7 whole cell lysatesLane 2: rat thymus tissue lysatesLane 3: rat RH-35 whole cell lysatesLane 4: mouse thymus tissue 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- MCM5 antigen affinity purified monoclonal antibody (Catalog # MBS1754026) 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 MCM5 at approximately 97KD. The expected band size for MCM5 is at 97KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of MCM5 using anti-MCM5 antibody (MBS1754026).MCM5 was detected in paraffin-embedded section of human gallbladder adenocarcinoma 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 2μg/ml mouse anti-MCM5 Antibody (MBS1754026) 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 2. IHC analysis of MCM5 using anti-MCM5 antibody (MBS1754026).MCM5 was detected in paraffin-embedded section of human gallbladder adenocarcinoma 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 2μg/ml mouse anti-MCM5 Antibody (MBS1754026) 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 MCM5 using anti-MCM5 antibody (MBS1754026).MCM5 was detected in paraffin-embedded section of human lung 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 2μg/ml mouse anti-MCM5 Antibody (MBS1754026) 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 MCM5 using anti-MCM5 antibody (MBS1754026).MCM5 was detected in paraffin-embedded section of human lung 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 2μg/ml mouse anti-MCM5 Antibody (MBS1754026) 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 MCM5 using anti-MCM5 antibody (MBS1754026).MCM5 was detected in paraffin-embedded section of human pancreatic 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 2μg/ml mouse anti-MCM5 Antibody (MBS1754026) 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 MCM5 using anti-MCM5 antibody (MBS1754026).MCM5 was detected in paraffin-embedded section of human pancreatic 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 2μg/ml mouse anti-MCM5 Antibody (MBS1754026) 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 MCM5 using anti- MCM5 antibody (MBS1754026).MCM5 was detected in immunocytochemical section of A431 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 5μg/mL mouse anti- MCM5 Antibody (MBS1754026) 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 MCM5 using anti- MCM5 antibody (MBS1754026).MCM5 was detected in immunocytochemical section of A431 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 5μg/mL mouse anti- MCM5 Antibody (MBS1754026) 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 A549 cells using anti-MCM5 antibody (MBS1754026).Overlay histogram showing A549 cells stained with MBS1754026 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- MCM5 Antibody (MBS1754026, 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 A549 cells using anti-MCM5 antibody (MBS1754026).Overlay histogram showing A549 cells stained with MBS1754026 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- MCM5 Antibody (MBS1754026, 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. )
2. Tsuruga, H., Yabuta, N., Hashizume, K., Ikeda, M., Endo, Y., Nojima, H. Expression, nuclear localization and interactions of human MCM/P1 proteins. Biochem. Biophys. Res. Commun. 236: 118-125, 1997.
3. Vetro, A., Savasta, S., Russo Raucci, A., Cerqua, C., Sartori, G., Limongelli, I., Forlino, A., Maruelli, S., Perucca, P., Vergani, D., Mazzini, G., Mattevi, A., Stivala, L. A., Salviati, L., Zuffardi, O. MCM5: a new actor in the link between DNA replication and Meier-Gorlin syndrome. Europ. J. Hum. Genet. 25: 646-650, 2017.
NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene is structurally very similar to the CDC46 protein from S. cerevisiae, a protein involved in the initiation of DNA replication. The encoded protein is a member of the MCM family of chromatin-binding proteins and can interact with at least two other members of this family. The encoded protein is upregulated in the transition from the G0 to G1/S phase of the cell cycle and may actively participate in cell cycle regulation. [provided by RefSeq, Jul 2008]
Uniprot Description
MCM5: 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. Interacts with MCMBP. Belongs to the MCM family.
Protein type: EC 3.6.4.12; Cell cycle regulation
Chromosomal Location of Human Ortholog: 22q13.1
Cellular Component: nucleoplasm; MCM complex; membrane; nucleus
Molecular Function: protein binding; DNA helicase activity; DNA binding; ATP binding
Biological Process: DNA replication initiation; mitotic cell cycle; DNA strand elongation during DNA replication; DNA replication; DNA duplex unwinding; G1/S transition of mitotic cell cycle
Research Articles on MCM5
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Product Notes
The MCM5 mcm5 (Catalog #AAA1754026) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-MCM5 Antibody (monoclonal, 4G10) reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's MCM5 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: 0.25-0.5ug/ml|Human, Mouse, Rat| IHC-P: 2-5ug/ml|Human| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the MCM5 mcm5 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 "MCM5, 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.Item has been added to Shopping Cart
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