HCLS1 抗体
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北京 101111
Quick Overview for HCLS1 抗体 (ABIN5541285)
抗原
See all HCLS1 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 HS1
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纯化方法
- Protein A agarose
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免疫原
- Recombinant human HS1
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亚型
- IgG1
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anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (AA 266-355) antibody
HCLS1 适用: 人 WB, ELISA, IHC (p), PLA, RNAi 宿主: 小鼠 Monoclonal 1A8 unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (N-Term) antibodyHCLS1 适用: 人, 小鼠, 大鼠, Cow, 犬, 马, 兔, 豚鼠 WB, IHC 宿主: 兔 Polyclonal unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (AA 266-355) antibodyHCLS1 适用: 人 WB, ELISA, IHC (p), IF 宿主: 小鼠 Monoclonal 2A9 unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (AA 161-260) antibodyHCLS1 适用: 小鼠 WB, ELISA, IHC (p), IF (cc), IF (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (AA 1-280) antibodyHCLS1 适用: 人 WB, IHC 宿主: 兔 Polyclonal unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (AA 266-356) antibodyHCLS1 适用: 人 WB, ELISA, IHC, IHC (p) 宿主: 小鼠 Monoclonal 2A8 unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (C-Term) antibodyHCLS1 适用: 人 WB, IHC (p) 宿主: 兔 Polyclonal unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (AA 28-77) antibodyHCLS1 适用: 人, 小鼠, 大鼠, Cow, 犬, 马, 兔, 豚鼠, Pig WB, IHC, IHC (p) 宿主: 兔 Polyclonal unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (AA 362-391), (C-Term) antibodyHCLS1 适用: 人 WB, IHC (p) 宿主: 兔 Polyclonal RB34076 unconjugated
anti-Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1) (C-Term) antibodyHCLS1 适用: 人 WB, ELISA 宿主: 兔 Polyclonal unconjugated
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应用备注
- Western blot: 1 μg/mL for chemiluminescence detection system. Immunoprecipitation: 10 μg/500 μL of cell extract from 5x10 6 cells.
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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 protease inhibitors at appropriate concentrations. Incubate it at 4 o C with rotating for 30 minutes thereafter, briefly sonicate the mixture (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 sample per lane on a 1-mm-thick SDS-polyacrylamide gel and carry out 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 manufacturer's manual for precise transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 5 % 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 suggested in the APPLICATIONS for 1 hour at room temperature. (The concentration of antibody will depend on the conditions.) 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 off the membrane, and incubate membrane with an appr opriate chemiluminescence reagent for 1 minute. 12) Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 13) Expose the membrane onto an X-ray film in a dark room for 3 minutes. 14) Develop the film under usual settings. The conditions for exposure and development may vary (Positive controls for Western blotting Raji, Jurkat, WR19L, PC12) Immunoprecipitation 1) Wash cells (approximately 1 x 10e7 cells) 3 times with PBS and resuspend them in 1000 μ L of cold Lysis buffer (50 mM Tris-HCl pH 7.2, 250 mM NaCl, 0.1 % NP-40, 2 mM EDTA, 10 % glycerol) containing protease inhibitors at appropriate concentrations. Incubate it at 4 o C with rotating for 30 minutes thereafter, briefly sonicate the mixture (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 fresh tube. 3) Add 50 μ L of 50 % protein A agarose beads in the supernatant. Incubate it at 4 o C with rotating for 60 minutes. 4) Centrifuge the tube at 12,000 x g for 5 minutes at 4 o C. Supernatant is equally divided into another fresh two tube. 5) Add the mouse IgG1 isotype control antibody or anti-HS1 (3A3) antibody at the amount of as suggested in the APPLICATIONS to the supernatant. Vortex briefly and incubate with gently agitation for 60-120 minutes at 4 o C. 6) Add 20 μ L of 50 % protein A agarose beads into the tube. Mix well and incubate with gentle agitation for 30-60 minutes at 4 o C. 7) Wash the beads 3-5 times with ice-cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds). 8) Resuspend the beads in 30 μ L of Laemmli's sample buffer, boil for 3-5 minutes, and centrifuge for 5 minutes. Use 15 μ L/lane for the SDS-PAGE analysis. (See SDS-PAGE & Western blotting .) (Positive control for Immunoprecipitation 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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- HCLS1 (Hematopoietic Cell-Specific Lyn Substrate 1 (HCLS1))
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别名
- hcls1
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背景
- The HS1 protein is one of the major substrates of non-receptor-type protein-tyrosine kinases and is phosphorylated immediately after crosslinking of the surface IgM on B cells. It is reported that HS1 protein plays a crucial role in the B-cell antigen receptor-mediated signal transduction pathway that leads to apoptosis.
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UniProt
- P14317
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途径
- Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Regulation of Actin Filament Polymerization, Myometrial Relaxation and Contraction, Maintenance of Protein Location
抗原
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