CDK6 抗体
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北京 101111
Quick Overview for CDK6 抗体 (ABIN487480)
抗原
See all CDK6 抗体适用
宿主
克隆类型
标记
应用范围
克隆位点
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特异性
- This antibody react with Human Cdk6 on Immunoprecipitation, Immunohistochemistry and Immunocytochemistry. It is reported that DCS-130 recognizes the C-terminus of Human Cdk6 (300-326) (See Reference 3).
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交叉反应 （详细）
- Species reactivity (tested):Human.
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产品特性
- Synonyms: PLSTIRE, Cell division protein kinase 6, Serine/threonine-protein kinase PLSTIRE
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纯化方法
- Protein-A Sepharose Chromatography.
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免疫原
- Recombinant Human Cdk6. Remarks: Hybridoma was established by fusion of mouse myeloma cell NS-2 with Balb/cmouse splenocyte.
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亚型
- IgG2a
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anti-Cyclin-Dependent Kinase 6 (CDK6) (AA 3-99) antibody
CDK6 适用: 人 WB, ELISA, IF, IHC (p), PLA 宿主: 小鼠 Monoclonal 8H4 unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) antibodyKO Validated CDK6 适用: 人 WB, IF, IHC (p), ICC 宿主: 兔 Polyclonal unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) antibodyCDK6 适用: 人 WB, IHC, ELISA 宿主: 兔 Polyclonal unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) antibodyCDK6 适用: 人, 大鼠 WB, IF, IHC (p), EIA 宿主: 小鼠 Monoclonal 8H4 unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) (AA 21-110) antibodyCDK6 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC (p), IF (cc), IF (p), FACS, IHC (fro) 宿主: 兔 Polyclonal unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) antibodyCDK6 适用: 人, 小鼠, 大鼠 WB 宿主: 兔 Monoclonal 4F7 unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) (pTyr24) antibodyCDK6 适用: 人 WB, IHC, ELISA, IF 宿主: 兔 Polyclonal unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) (N-Term) antibodyCDK6 适用: 人, 小鼠, 大鼠 WB, IHC, ELISA 宿主: 兔 Polyclonal unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) (AA 102-326) antibodyCDK6 适用: 人 IHC, ELISA, IF 宿主: 兔 Polyclonal unconjugated
anti-Cyclin-Dependent Kinase 6 (CDK6) antibodyCDK6 适用: 人 WB, ELISA, IP 宿主: 兔 Monoclonal 8G3 unconjugated Recombinant Antibody
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应用备注
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Immunocytochemistry: 10 μg/mLImmunoprecipitation: 10 μg/400 μL of cell extract from 5x10^6 cells. Positive Controls: HeLa, Jurkat cells. Immunohistochemistry: 10 μg/mLHeat treatment is necessary for Paraffin Embedded Sections. Microwave oven: 2 times for 10 minutes each 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. -
实验流程
- Immunoprecipitation1) 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 min at 4°C and transfer the supernatant toanother tube. 3) Add primary antibody as suggest in the APPLICATIONS into 400 µL of the supernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4oC. Add 20 ìL of 50%protein A Agarose resuspended in the cold Lysis buffer. Mix well and incubate with gentleagitation for 60 minutes at 4°C. 4) Wash the beads 3-5 times with the ice-cold Lysis buffer (centrifuge the tube at 2,500 x gfor 10 seconds). 5) Resuspend the beads in 20 μL of Laemmli’s sample buffer, boil for 3-5 min, andcentrifuge for 5 minutes. Use 10 μL/lane for the SDS-PAGE analysis. 6) Load 10 µL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel forelectrophoresis. 7) 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. 8) 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. 9) Incubate the membrane with 1 µg/mL of anti-Cdk6 as primary antibody diluted with PBS,pH 7. 2 containing 1% skimmed milk for 1 hour at room temperature. (The concentration ofantibody will depend on condition. )10) Wash the membrane with PBS-T [0. 05% Tween-20 in PBS] (5 minutes x 3 times). 11) 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. 12) Wash the membrane with PBS-T (5 minutes x 6 times). 13) Wipe excess buffer on the membrane, then incubate it with appropriatechemiluminescence reagent for 1 minute. Remove extra reagent from the membrane bydabbing with paper towel, and seal it in plastic wrap. 14) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. Thecondition for exposure and development may varyPositive Control for Immunoprecipitation: HeLa, Jurkat. Immunohistochemical 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: Place the slides put on staining basket in 500 mL beakerwith 500 mL of 10 mM citrate buffer (pH 6. 5). Cover the beaker with plastic wrap, then
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限制
- 仅限研究用
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浓度
- 1.0 mg/mL
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缓冲液
- PBS, pH 7.2 containing 50 % Glycerol without preservatives.
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储存液
- Without preservative
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储存条件
- -20 °C
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储存方法
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Store the antibody undiluted at -20 °C.
Shelf life: one year from despatch. -
有效期
- 12 months
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- CDK6 (Cyclin-Dependent Kinase 6 (CDK6))
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别名
- CDK6
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背景
- Cyclin-dependent kinase 6 (Cdk6) is a 38-40 kDa member of the Cdk family of mitotic kinases involved in cell cycle progression. The Cdk6 phosphoprotein, like Cdk4, associates with D-cyclins to regulate cell proliferation. Following translocation to the nucleus, the Cdk6 is phosphorylated by Cdk-Activating Kinase (CAK), and the Cdk6 complex, in association with PCNA, phosphorylates the retinoblastoma (Rb) protein. Overexpression of Cdk6 has been associated with several different types of cancer.Synonyms: Cell division protein kinase 6, PLSTIRE, Serine/threonine-protein kinase PLSTIRE
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基因ID
- 1021
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UniProt
- Q00534
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途径
- Cell Division Cycle, Mitotic G1-G1/S Phases
抗原
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