CHEK1 抗体
Quick Overview for CHEK1 抗体 (ABIN487487)
抗原
See all CHEK1 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 reacts with CHK1 (55 kDa) on Western blots.
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交叉反应 （详细）
- Species reactivity (tested):Human.
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产品特性
- Synonyms: CHEK1, CHEK-1, Serine/threonine-protein kinase Chk1, CHK1 checkpoint homolog
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纯化方法
- Protein-A Sepharose Chromatography.
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免疫原
- Human recombinant full length CHK1. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte
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亚型
- IgG2b
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anti-Checkpoint Kinase 1 (CHEK1) (pSer317) antibody
CHEK1 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-Checkpoint Kinase 1 (CHEK1) (pSer301) antibodyCHEK1 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-Checkpoint Kinase 1 (CHEK1) (pSer296) antibodyCHEK1 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-Checkpoint Kinase 1 (CHEK1) (pSer286) antibodyCHEK1 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-Checkpoint Kinase 1 (CHEK1) (pSer345) antibodyCHEK1 适用: 人 WB, IF, IHC (p), ICC 宿主: 兔 Polyclonal unconjugated
anti-Checkpoint Kinase 1 (CHEK1) (Middle Region) antibodyCHEK1 适用: 人, 小鼠, 大鼠, 犬, 兔, Cow, 马, 豚鼠 WB, IHC 宿主: 兔 Polyclonal unconjugated
anti-Checkpoint Kinase 1 (CHEK1) (pSer317) antibodyCHEK1 适用: 小鼠, 小鸡 ELISA, IHC (p), ICC, IF (cc), IF (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated
anti-Checkpoint Kinase 1 (CHEK1) antibodyCHEK1 适用: 人 WB, ELISA, IHC, IF 宿主: 兔 Monoclonal 2F2 unconjugated Recombinant Antibody
anti-Checkpoint Kinase 1 (CHEK1) antibodyCHEK1 适用: 人, 小鼠 WB, ELISA 宿主: 小鼠 Monoclonal 2G1D5 unconjugated
anti-Checkpoint Kinase 1 (CHEK1) (pSer317) antibodyCHEK1 适用: 人 WB, IHC (p), DB 宿主: 兔 Polyclonal RB7845-RB18425 unconjugated
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应用备注
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Western Blot: 1 μg/mL mL for chemiluminescence detection system. Positive Controls: HeLa, MCF7, Raji Cells. Immunoprecipitation: 2 μg/200 μL of cell extract from 5x10^6 cells. Positive Control: Raji. 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 Tissue. 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
- 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: HeLa, MCF7, RajiImmunoprecipitation1) 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. 3) Add primary antibody as suggested in the APPLICATIONS into 200 μL of the supernatant. Mix well and incubate with gentle agitation for 30-120minutes at 4°C. Add 20 μL of 50% protein A agarose beads resuspended in the cold Lysisbuffer. Mix well and incubate with gentle agitation for 60 minutes at 4°C. 4) Wash the beads 3-5 times with ice-cold Lysis buffer (centrifuge the tube at 2,500 x g for10 seconds). 5) Resuspend the beads in 20 μL of Laemmli’s sample buffer, boil for 3-5 minutes, andcentrifuge for 5 minutes. Use 10 μL/lane for the SDS-PAGE analysis. (See SDS-PAGE & Western blotting. )Positive Controls for immunoprecipitation: Raji.
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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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- CHEK1 (Checkpoint Kinase 1 (CHEK1))
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别名
- CHK1
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背景
- The DNA damage checkpoint is a signal transduction pathway that delays entry into mitosis following DNA damage. When DNA is damaged, Chk1 acts downstream of ATM to elicit appropriate responses such as cell cycle arrest. When activated by ATM, Chk1 phosphorylates serines 123, 178, 278, and 292 of the S phase-promoting CDC25A phosphatase, which accelerates IR (ionizing radiation)-induced degradation of CDC25A.Synonyms: CHEK-1, CHEK1, CHK1 checkpoint homolog, Serine/threonine-protein kinase Chk1
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基因ID
- 1111
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UniProt
- O14757
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途径
- p53 Pathway, Apoptosis, Cell Division Cycle, DNA Damage Repair
抗原
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