Western Blot (WB)
(Figure 1. Western blot analysis of MCM4 using anti-MCM4 antibody (MBS1753514).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 U20S whole cell lysatesLane 3: human MCF-7 whole cell lysatesLane 4: human HEK293 whole cell lysatesLane 5: human Jurkat whole cell lysatesLane 6: human HL-60 whole cell lysatesLane 7: rat liver tissue lysatesLane 8: mouse NIH/3T3 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 rabbit anti-MCM4 antigen affinity purified polyclonal antibody (Catalog # MBS1753514) at 0. 25 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for MCM4 at approximately 97KD. The expected band size for MCM4 is at 97KD. )
Rabbit MCM4 Polyclonal Antibody | anti-MCM4 antibody
Anti-MCM4 Antibody
IHC-P: 2-5ug/ml|Human, Mouse, Rat|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of MCM4 using anti-MCM4 antibody (MBS1753514).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 U20S whole cell lysatesLane 3: human MCF-7 whole cell lysatesLane 4: human HEK293 whole cell lysatesLane 5: human Jurkat whole cell lysatesLane 6: human HL-60 whole cell lysatesLane 7: rat liver tissue lysatesLane 8: mouse NIH/3T3 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 rabbit anti-MCM4 antigen affinity purified polyclonal antibody (Catalog # MBS1753514) at 0. 25 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for MCM4 at approximately 97KD. The expected band size for MCM4 is at 97KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of MCM4 using anti-MCM4 antibody (MBS1753514).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 U20S whole cell lysatesLane 3: human MCF-7 whole cell lysatesLane 4: human HEK293 whole cell lysatesLane 5: human Jurkat whole cell lysatesLane 6: human HL-60 whole cell lysatesLane 7: rat liver tissue lysatesLane 8: mouse NIH/3T3 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 rabbit anti-MCM4 antigen affinity purified polyclonal antibody (Catalog # MBS1753514) at 0. 25 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for MCM4 at approximately 97KD. The expected band size for MCM4 is at 97KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 was detected in paraffin-embedded section of mouse colon 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 was detected in paraffin-embedded section of mouse colon 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 was detected in paraffin-embedded section of rat colon 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 was detected in paraffin-embedded section of rat colon 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 was detected in paraffin-embedded section of human breast 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 was detected in paraffin-embedded section of human breast 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 6. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 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 2μg/ml rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 6. IHC analysis of MCM4 using anti-MCM4 antibody (MBS1753514).MCM4 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 2μg/ml rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 # MBS176451) with DAB as the chromogen. )
Immunofluorescence (IF)
(Figure 7. IF analysis of MCM4 using anti- MCM4 antibody (MBS1753514).MCM4 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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 7. IF analysis of MCM4 using anti- MCM4 antibody (MBS1753514).MCM4 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 rabbit anti-MCM4 Antibody (MBS1753514) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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 8. Flow Cytometry analysis of A549 cells using anti-MCM4 antibody (MBS1753514).Overlay histogram showing A549 cells stained with MBS1753514 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MCM4 Antibody (MBS1753514, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 8. Flow Cytometry analysis of A549 cells using anti-MCM4 antibody (MBS1753514).Overlay histogram showing A549 cells stained with MBS1753514 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MCM4 Antibody (MBS1753514, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Groth, A., Corpet, A., Cook, A. J. L., Roche, D., Bartek, J., Lukas, J., Almouzni, G. Regulation of replication fork progression through histone supply and demand. Science 318: 1928-1931, 2007.
3. Hu B, Burkhart R, Schulte D et al (1994). "The P1 family: a new class of nuclear mammalian proteins related to the yeast Mcm replication proteins". Nucleic Acids Res. 21 (23): 5289-93.
NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene is one of the highly conserved mini-chromosome maintenance proteins (MCM) that are essential for 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. The MCM complex consisting of this protein and MCM2, 6 and 7 proteins possesses DNA helicase activity, and may act as a DNA unwinding enzyme. The phosphorylation of this protein by CDC2 kinase reduces the DNA helicase activity and chromatin binding of the MCM complex. This gene is mapped to a region on the chromosome 8 head-to-head next to the PRKDC/DNA-PK, a DNA-activated protein kinase involved in the repair of DNA double-strand breaks. Alternatively spliced transcript variants encoding the same protein have been reported. [provided by RefSeq, Jul 2008]
Uniprot Description
MCM4: a mini-chromosome maintenance protein, essential for the initiation of eukaryotic genome replication. The MCM hexameric protein complex is a key component of the pre-replication complex. The complex consisting of this protein and MCM2, 6 and 7 possesses DNA helicase activity. Phosphorylation by CDC2 reduces the DNA helicase activity and chromatin binding of the complex.
Protein type: EC 3.6.4.12; DNA replication
Chromosomal Location of Human Ortholog: 8q11.2
Cellular Component: nucleoplasm; membrane; MCM complex; nucleus
Molecular Function: ATP-dependent DNA helicase activity; protein binding; single-stranded DNA binding; ATP binding
Biological Process: DNA unwinding during replication; DNA replication initiation; DNA strand elongation during DNA replication; mitotic cell cycle; DNA replication; G1/S transition of mitotic cell cycle
Disease: Natural Killer Cell And Glucocorticoid Deficiency With Dna Repair Defect
Research Articles on MCM4
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Product Notes
The MCM4 mcm4 (Catalog #AAA1753514) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-MCM4 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's MCM4 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), Direct ELISA (EIA). WB: 0.1-0.25ug/ml|Human, Mouse, Rat| IHC-P: 2-5ug/ml|Human, Mouse, Rat| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the MCM4 mcm4 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 "MCM4, Polyclonal 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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