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|
FCM (Flow Cytometry)
(Figure 6. Flow Cytometry analysis of A549 cells using anti-MCM5 antibody (AAA19377).Overlay histogram showing A549 cells stained with AAA19377 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- MCM5 Antibody (AAA19377, 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.)
IF (Immunofluorescence)
(Figure 5. IF analysis of MCM5 using anti- MCM5 antibody (AAA19377).MCM5 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- MCM5 Antibody (AAA19377) 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.)
IHC (Immunohistochemistry)
(Figure 4. IHC analysis of MCM5 using anti-MCM5 antibody (AAA19377).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 (AAA19377) 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 3. IHC analysis of MCM5 using anti-MCM5 antibody (AAA19377).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 (AAA19377) 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 2. IHC analysis of MCM5 using anti-MCM5 antibody (AAA19377).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 (AAA19377) 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 # with DAB as the chromogen.)
WB (Western Blot)
(Figure 1. Western blot analysis of MCM5 using anti-MCM5 antibody (AAA19377).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 # AAA19377) 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 # with Tanon 5200 system. A specific band was detected for MCM5 at approximately 97KD. The expected band size for MCM5 is at 97KD.)
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.
