MCM7 Antikörper

Produktnummer ABIN487308

Dieses Anti-MCM7-Antikörper ist ein Maus Monoklonal-Antikörper zur Detektion von MCM7 in WB und IHC (p). Geeignet für Human und Maus.

Kurzübersicht für MCM7 Antikörper (ABIN487308)

Target: MCM7 (Minichromosome Maintenance Complex Component 7 (MCM7))

Reaktivität: Human, Maus

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser MCM7 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Klon: DCS-141

Produktdetails anti-MCM7 Antikörper

Spezifität: This antibody reacts with MCM7 (90 kDa) on Western blotting.

Kreuzreaktivität (Details): Species reactivity (tested):Human and Mouse.

Produktmerkmale: Synonyms: CDC47, MCM2, CDC47 homolog, P1.1-MCM3, minichromosome maintenance complexcomponent 7, DNA replication licensing factor MCM7

Aufreinigung: Protein-A Sepharose Chromatography.

Immunogen: Human recombinant full MCM7. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte.

Isotyp: IgG1

Anwendungsinformationen

Applikationshinweise: Western Blot: 1 μg/mL for chemiluminescence detection system. Positive Controls: HeLa, A-431, Jurkat, C2C12 cells. Immunohistochemistry: 1 μg/mLHeat treatment is necessary for Paraffin Embedded Sections. Microwave oven: 2 times for 10 minutes each in citrate buffer ( pH 6.5). Positive Control: Tonsil. Detailed procedure is provided in Protocols.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protokoll: SDS-PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. Measure the protein concentration of the supernatant and add the Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli’s sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 μL of the sample per lane in a 1mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for the transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at room temperature, or overnight at 4°C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7. 2 containing 1%skimmed milk as suggested in the APPLICATIONS for 1 hour at room temperature. (Theoptimal antibody concentration will depend on the experimental conditions. 8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1: 10000 POD-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 10) Wash the membrane with PBS (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriatechemiluminescence reagents for 1 minute. Remove extra reagent from the membrane bydabbing with a paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. Theconditions for exposure and development may vary. Positive Controls for Western blotting: Jurkat, HeLa, A-431, C2C12Immunohistochemical Staining for Paraffin-Embedded Sections: SAB method1) Deparaffinize the sections with Xylene 3 times for 3-5 minutes each. 2) Wash the slides with Ethanol 3 times for 3-5 minutes each. 3) Wash the slides with PBS 3 times for 3-5 minutes each. 4) Heat treatmentHeat treatment by microwave oven: Place the slides put on staining basket in 500 mLbeaker with 500 mL citrate buffer (pH 6. 5). Cover the beaker with plastic wrap, thenprocess the slides 2 times for 10 minutes each at 500 W with microwave oven. Let theslides cool down in the beaker at room temperature for about 40 minutes. 5) Remove the slides from the citrate buffer and cover each section with 3% H2O2 for 10minutes at room temperature to block endogenous peroxidase activity. Wash 3 times inPBS for 5 minutes each. 6) Remove the slides from PBS, wipe gently around each section and cover tissues withProtein Blocking Agent for 5 minutes to block non-specific antibody staining. Do not wash. 7) Tip off the blocking buffer, wipe gently around each section and cover tissues withprimary antibody diluted with PBS containing 1% BSA as suggested in the APPLICATIONS.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Konzentration: 1.0 mg/mL

Buffer: PBS, pH 7.2 containing 50 % Glycerol without preservatives.

Konservierungsmittel: Without preservative

Lagerung: -20 °C

Informationen zur Lagerung: Store the antibody undiluted at -20 °C.
Shelf life: one year from despatch.

Haltbarkeit: 12 months

Details zu MCM7

Target: MCM7 (Minichromosome Maintenance Complex Component 7 (MCM7))

Andere Bezeichnung: MCM7

Hintergrund: MCM7, also known as CDC47, is a member of the MCM (multicopy maintenance) protein family which act as part of the regulatory machinery allowing DNA to replicate only once during S phase. In quiescent cells, human MCM7 mRNA is almost undetectable. Stimulation of cells to enter the cell cycle results in an increase in MCM7 protein during late G1 to S phase, making it a good marker for proliferation. MCM7 proteins are located in nuclei of the proliferative components of normal lymph nodes, bone marrow, epidermis and mucosa, where they exist both free in the nucleoplasm and bound to chromatin. Malignant tumors expressed increased levels of MCM7, suggesting MCM7 may play a role in normal and neoplastic cell growth in vivoSynonyms: CDC47, CDC47 homolog, DNA replication licensing factor MCM7, MCM2, P1.1-MCM3, minichromosome maintenance complex component 7

Gen-ID: 4176

UniProt: P33993

Pathways: DNA Reparatur, Mitotic G1-G1/S Phases, DNA Replication, Chromatin Binding, Synthesis of DNA