CDC27 antibody (C-Term)

Catalog No. ABIN492595

Product Details anti-CDC27 Antibody

Target: CDC27 (Cell Division Cycle 27 Homolog (S. Cerevisiae) (CDC27))

Binding Specificity: AA 814-823, C-Term

Reactivity: Human, Rat, Mouse, Dog, Cow, Xenopus laevis

Host: Mouse

Clonality: Monoclonal

Conjugate: This CDC27 antibody is un-conjugated

Application: Western Blotting (WB), Immunoprecipitation (IP), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Enzyme Immunoassay (EIA)

Specificity: This antibody reacts with 97 kDa membrane protein CDC27 on Western blotting, Immunoprecipitation and Immunohistochemistry.

Cross-Reactivity (Details): Species reactivity (expected):Bovine, Canine and Xenopus leavis.
Species reactivity (tested):Human, Mouse, Rat.

Characteristics: Synonyms: CDC27Hs, H-NUC, D0S1430E, D17S978E, Cell division cycle protein 27 homolog

Purification: Protein-A Agarose Chromatography of hybridoma supernatant.

Immunogen: Peptide corresponding to C-terminal of Human CDC27 (aa 814-823).

Clone: AF3-1

Isotype: IgG2b

Application Details

Application Notes: Western blotting: 1 μg/mL for chemiluminescence detection system. Positive Controls: Jurkat, Raji, K562, HL-60. Immunoprecipitation: 2 μg/200 μL of cell extract. Positive Control: Raji. Immunohistochemistry on Paraffin Embedded Sections: 10 μg/mL. (Heat treatment is necessary)Autoclave: 10 minutes at 110 °C in 10 mM 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.

Protocol: 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 cold Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli's sample buffer. 4) Boil the samples for 3 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 precise 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 suggest in the APPLICATIONS for 1 hour at room temperature. (Theconcentration of antibody will depend on condition. )8) Wash the membrane with PBS-T [0. 05% Tween-20 in PBS] (5 minutes x 3 times). 9) Incubate the membrane with the 1: 10,000 HRP-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at RT. 10) Wash the membrane with PBS-T (10 minutes x 3 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriatechemiluminescence reagent for 1 minute. 12) Remove extra reagent from the membrane by dabbing with paper towel, and seal it inplastic wrap. 13) Expose to an X-ray film in a dark room for 3 minutes. 14) Develop the film as usual. The condition for exposure and development may vary. Positive Controls: Jurkat, Raji, K562, HL-60, MCF-7Immunoprecipitation1) Wash the cells 3 x 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. 3) Add primary antibody as suggested in the APPLICATIONS into 200 µL of the supernatant.

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

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

Preservative: Without preservative

Storage: -20 °C

Storage Comment: Store the antibody (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch.

Expiry Date: 12 months

Target Details for CDC27

Target: CDC27 (Cell Division Cycle 27 Homolog (S. Cerevisiae) (CDC27))

Alternative Name: CDC27 (CDC27 Products)

Synonyms: ANAPC3 antibody, APC3 antibody, CDC27Hs antibody, D0S1430E antibody, D17S978E antibody, HNUC antibody, NUC2 antibody, wu:fb54g06 antibody, wu:fc88c12 antibody, AI452358 antibody, BC023187 antibody, CDC27 antibody, NV10210 antibody, cell division cycle 27 antibody, cell division cycle 27 S homeolog antibody, cell division cycle protein 27 homolog antibody, anaphase-promoting complex subunit Apc3 antibody, CDC27 antibody, cdc27 antibody, Cdc27 antibody, cdc27.S antibody, LOC100123129 antibody, nuc2 antibody

Background: CDC16, CDC27 and CDC34 are components of the APC (anaphase-promoting complex) that also play a role as components of the ubiquitin- conjugating E3 enzyme. A 20S complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B, resulting in cyclin B/Cdk complex degradation. CDC27 is therefore required for progression of the cell cycle, where it executes essential mitotic functions near the metaphase/anaphase transition. CDC27 is ~97 kDa and is localized to the centrosome and mitotic spindle.Synonyms: CDC27Hs, Cell division cycle protein 27 homolog, D0S1430E, D17S978E, H-NUC

Gene ID: 996

UniProt: P30260