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Details for Product No. ABIN968618

Minichromosome Maintenance Complex Component 6 (MCM6) (AA 670-792) antibody

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Antigen
Synonyms Mis5, P105MCM, MCG40308, Mcmd6, ASP-l1, D1Wsu22e, CG4039, DmMCM6, DmeMCM6, Dmel\\CG4039, MCM6, McM6, fs(1)K1214, mcm6
Epitope
»Alternatives AA 670-792
Reactivity
»Alternatives Human
Host
»Alternatives Mouse
Clonality (Clone) Monoclonal ()
Application
»Alternatives Western Blotting (WB), BioImaging (BI)
Pubmed 4 references available
Catalog no. ABIN968618
Quantity 50 µg
Price
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Immunogen Human MCM6
Clone 1
Isotype IgG1
Characteristics 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Purity Purified
Alternative Name MCM6
Background Mcm proteins are nuclear proteins that perform essential functions in the regulation of chromatin replication. They are primarily bound to chromatin during G1 and the beginning of S-phase, but are found in the nucleosol in postreplicative cells. Release of the bound Mcm proteins may be a mechanism for prevention of chromatin re-replication during S-phase. MCM6 is homologous to the yeast replication protein Mis5 and contains a DEFD-box motif and nucleotide-binding domain in the central region that are conserved in Mcm proteins. MCM6 forms a complex with MCM2, -4, and -7, which binds histone H3. In addition, the subcomplex of MCM4, -6, and -7 has helicase activity, which is mediated by the ATP-binding activity of MCM6 and the DNA-binding activity of MCM4. In Xenopus, formation of pre-replication complexes that include Mcm protein complexes requires phosphorylation, since intermediate phosphorylation of MCM4 correlates with Mcm protein complex formation and chromatin binding. Thus, Mcm proteins form heterocomplexes that participate in many different functions required for chromatin replication, such as DNA binding, helicase activity, and chromatin structure rearrangement.
Molecular Weight 105 kDa
Research Area Chromatin and Nuclear Signaling, Chromatin Binding Proteins, DNA/RNA
Application Notes Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.
Comment

Related Products: ABIN968535

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
Preservative Sodium azide
Storage -20 °C
Product cited in: Holthoff, Hameister, Knippers: "A novel human Mcm protein: homology to the yeast replication protein Mis5 and chromosomal location." in: Genomics, Vol. 37, Issue 1, pp. 131-4, 1997 (PubMed).

You, Komamura, Ishimi: "Biochemical analysis of the intrinsic Mcm4-Mcm6-mcm7 DNA helicase activity." in: Molecular and cellular biology, Vol. 19, Issue 12, pp. 8003-15, 2000 (PubMed).

Maiorano, Lemaître, Méchali: "Stepwise regulated chromatin assembly of MCM2-7 proteins." in: The Journal of biological chemistry, Vol. 275, Issue 12, pp. 8426-31, 2000 (PubMed).

Pereverzeva, Whitmire, Khan et al.: "Distinct phosphoisoforms of the Xenopus Mcm4 protein regulate the function of the Mcm complex." in: Molecular and cellular biology, Vol. 20, Issue 10, pp. 3667-76, 2000 (PubMed).

Alternatives for antigen "Minichromosome Maintenance Complex Component 6 (MCM6)", type "Antibodies"
Hosts (29), (3), (2)
Reactivities (31), (12), (7), (2), (1)
Applications (32), (15), (11), (4), (2), (2), (2), (1), (1), (1)
Epitopes (6), (4), (2), (2), (2), (1)
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