Product Info Summary
SKU: | A02301-1 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-MCM4 Antibody Picoband™
SKU/Catalog Number
A02301-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-MCM4 Antibody Picoband™ catalog # A02301-1. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
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.
Cite This Product
Anti-MCM4 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02301-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human MCM4 recombinant protein (Position: M361-A677).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A02301-1 is reactive to MCM4 in Human, Mouse, Rat
Applications
A02301-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
97 kDa
Calculated molecular weight
30392 MW
Background of MCM4
MCM4 (MINICHROMOSOME MAINTENANCE, S. CEREVISIAE, HOMOLOG OF, 4), also called CDC21, S. POMBE, HOMOLOG OF, is a protein that in humans is encoded by the MCM4 gene. MCM4 is one of the highly conserved mini-chromosome maintenance proteins (MCM) that are essential for the initiation of eukaryotic genome replication. The MCM4 gene is mapped to 8q11.21. The 864-amino acid MCM4 protein has an observed molecular mass of 97 kD by SDS-PAGE, which is similar to its calculated molecular mass of 96.6 kD. Western blot analysis with and without phosphatase treatment suggested that MCM4 is highly phosphorylated in mitotic cells. In the absence of DDK, CDK phosphorylation at the distal part of the Mcm4 NSD becomes crucial.MCM4 encodes a subunit of the MCM2-7 complex (also known as MCM2-MCM7), the replication licensing factor and presumptive replicative helicase.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Assay dilution & Images
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.25μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of MCM4 using anti-MCM4 antibody (A02301-1).
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 U20S whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human HEK293 whole cell lysates,
Lane 5: human Jurkat whole cell lysates,
Lane 6: human HL-60 whole cell lysates,
Lane 7: rat liver tissue lysates,
Lane 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 # A02301-1) at 0.25 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-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 # EK1002) with Tanon 5200 system. A specific band was detected for MCM4 at approximately 97KD. The expected band size for MCM4 is at 97KD.
Click image to see more details
Figure 2. IHC analysis of MCM4 using anti-MCM4 antibody (A02301-1).
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 (A02301-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of MCM4 using anti-MCM4 antibody (A02301-1).
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 (A02301-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of MCM4 using anti-MCM4 antibody (A02301-1).
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 (A02301-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of MCM4 using anti-MCM4 antibody (A02301-1).
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 (A02301-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of MCM4 using anti-MCM4 antibody (A02301-1).
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 (A02301-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 7. IF analysis of MCM4 using anti-MCM4 antibody (A02301-1).
MCM4 was detected in immunocytochemical section of A431 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 5μg/mL rabbit anti-MCM4 Antibody (A02301-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.
Click image to see more details
Figure 8. Flow Cytometry analysis of A549 cells using anti-MCM4 antibody (A02301-1).
Overlay histogram showing A549 cells stained with A02301-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MCM4 Antibody (A02301-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For MCM4 (Source: Uniprot.org, NCBI)
Gene Name
MCM4
Full Name
DNA replication licensing factor MCM4
Weight
30392 MW
Superfamily
MCM family
Alternative Names
CDC21 homolog; CDC21; CDC54; DNA replication licensing factor MCM4; EC 3.6.4.12; hCdc21; homolog of S. pombe cell devision cycle 21; MCM4 minichromosome maintenance deficient 4 (S. cerevisiae); MCM4 minichromosome maintenance deficient 4; MGC33310; minichromosome maintenance complex component 4; minichromosome maintenance deficient 4; P1-Cdc21 MCM4 CDC21, CDC54, IMD54, NKCD, NKGCD, P1-CDC21, hCdc21 minichromosome maintenance complex component 4 DNA replication licensing factor MCM4|homolog of S. pombe cell devision cycle 21|minichromosome maintenance deficient 4
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on MCM4, check out the MCM4 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for MCM4: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-MCM4 Antibody Picoband™ (A02301-1)
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