Separase 抗体 (AA 1866-1966)
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Quick Overview for Separase 抗体 (AA 1866-1966) (ABIN5540331)
抗原
See all Separase (ESPL1) 抗体适用
宿主
克隆类型
标记
应用范围
克隆位点
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抗原表位
- AA 1866-1966
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特异性
- This antibody reacts with human Separase (190 kDa).
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交叉反应 （详细）
- Does not work with Mouse (WR19L, U937, P19, NIH/3T3) and Hamster (CHO).
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纯化方法
- Protein A agarose
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免疫原
- Separase (1866-1966 aa)
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亚型
- IgG1
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anti-Extra Spindle Poles Like 1 (ESPL1) (AA 586-701) antibody
ESPL1 适用: 人 ELISA, WB, IHC (p) 宿主: 小鼠 Monoclonal 6H6 unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) antibodyESPL1 适用: 人, 小鼠 ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) (AA 586-702) antibodyESPL1 适用: 人 ELISA, IHC, WB, IHC (p) 宿主: 小鼠 Monoclonal 6H6 unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) (Tyr489) antibodyESPL1 适用: 人, 小鼠 ELISA, IHC, WB, IF 宿主: 兔 Polyclonal unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) (pSer1126) antibodyESPL1 适用: 人 IHC (p), DB 宿主: 兔 Polyclonal RB7943 unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) (pSer801) antibodyESPL1 适用: 人 IHC, WB, IF, IC 宿主: 兔 Polyclonal unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) (AA 767-816) antibodyESPL1 适用: 人, 小鼠 ELISA, IHC, WB, IF 宿主: 兔 Polyclonal unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) (pSer801) antibodyESPL1 适用: 人 ELISA, IHC, WB, IF 宿主: 兔 Polyclonal unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) (AA 1220-1500) antibodyESPL1 适用: 人 WB, IF 宿主: 兔 Polyclonal unconjugated
anti-Extra Spindle Poles Like 1 (ESPL1) (AA 1292-1495) antibodyESPL1 适用: 人 ELISA, IF 宿主: 兔 Polyclonal unconjugated
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应用备注
- Western blot: 1 μg/mL for chemiluminescence detection system. Not reconmmended for Immunoprecipitation, Immunohistochemistry, and Immunocytochemistry.
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实验流程
- SDS-PAGE & Western Blotting 1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1 % NP-40, 2 mM EDTA, 10 % glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the Lysis buffer 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 μ L of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in 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, pH 7.2) for 1 hour at room temperature, or overnight at 4 o 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 ro om temperature. (The concentration 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 with 1 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS-T (5 minutes x 6 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriate chemilumin escence reagent for 1 minute. Remove extra reagent from the membrane by dabbing with paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 10 minutes. Develop the film as usual. The condition for exposure and development may vary. (Positive controls for Western blotting HeLa, Raji, ZR-75-1, MCF-7, HL-60, Jurkat)
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限制
- 仅限研究用
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状态
- Liquid
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缓冲液
- PBS containing 50 % glycerol, pH 7.2. No preservative is contained.
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储存液
- Azide free
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储存条件
- -20 °C
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储存方法
- Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.
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- Separase (ESPL1) (Extra Spindle Poles Like 1 (ESPL1))
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别名
- separin,espl1
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背景
- Separase (190 kDa) is a cysteine protease that is essential for chromosome segregation. During cell division the sister chromatids are held together by the cohesin complex. At the onset of anaphase, separase cleaves the SCC1/RAD21 subunit of cohesin, allowing the chromatids to separate and divide with the cell. During most of the cell cycle, separase is inactivated by the securin/PDS-1 protein, which probably covers its active site. Separase is also down re gulated by self-cleavage to a ~65 kDa form. Separase can be used as a marker for centrosomes in mitotic cells.
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UniProt
- Q14674
抗原
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