CDC25C Antikörper

Produktnummer ABIN487484

Der Maus Monoklonal Anti-CDC25C-Antikörper wurde für WB validiert. Er ist geeignet, CDC25C in Proben von Human, Maus und Ratte zu detektieren.

Kurzübersicht für CDC25C Antikörper (ABIN487484)

Target: CDC25C (Cell Division Cycle 25 Homolog C (S. Pombe) (CDC25C))

Reaktivität: Human, Maus, Ratte

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser CDC25C Antikörper ist unkonjugiert

Applikation: Western Blotting (WB)

Klon: DCS-193

Produktdetails anti-CDC25C Antikörper

Spezifität: This antibody reacts with CDC25C (60 kDa) on Western blotting.

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

Produktmerkmale: Synonyms: M-phase inducer phosphatase 3, Dual specificity phosphatase Cdc25C, Cell Division Cycle25C

Aufreinigung: Protein-A Agarose Chromatography

Immunogen: Human CDC25C. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte.

Isotyp: IgG1

Anwendungsinformationen

Applikationshinweise: Western Blot: 10 μg/mL for chemiluminescence detection system. Positive Controls: HeLa, A431, ZR75-1, HEp-2, NIH/3T3, PC12. 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: 10,000 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: HeLa, A431, ZR75-1, HEp-2, NIH/3T3, PC12

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 CDC25C

Target: CDC25C (Cell Division Cycle 25 Homolog C (S. Pombe) (CDC25C))

Andere Bezeichnung: CDC25C

Hintergrund: The activity of cyclin-dependent kinases is regulated by their phosphorylation status, which is controlled by the antagonistic action of CDC25 phosphatases. Three CDC25 genes are present in human cells: CDC25A, CDC25B and CDC25C. CDC25C is a ~56 kDa dual specific protein phosphatase that controls entry into mitosis by regulating the dephosphorylation of the Cdk1/cyclin B complex. Phosphorylation of Cdc25C creates a binding site for 14-3-3 protein which inhibits Cdc25C. This prevents activation of the Cdk1/cyclin B complex and prevents mitotic entry. Cdc25C is phosphorylated by checkpoint kinases throughout interphase but becomes dephosphorylated during mitosis. Cdc25C is also regulated by p53 via a p53 response element in its promoter, and it is predominantly expressed in the G2 phase of the cell cycle.Synonyms: Cell Division Cycle 25C, Dual specificity phosphatase Cdc25C, M-phase inducer phosphatase 3

Gen-ID: 995

UniProt: P30307

Pathways: Zellzyklus, M Phase