TPT1 抗体 (AA 1-172)
Quick Overview for TPT1 抗体 (AA 1-172) (ABIN5541439)
抗原
See all TPT1 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 1-172
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特异性
- This antibody reacts with HRF (23 kDa).
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纯化方法
- Protein-A agarose
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免疫原
- Full length human recombinant GST-HRF (1-172 a.a.)
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亚型
- IgG1
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anti-Tumor Protein, Translationally-Controlled 1 (TPT1) (AA 35-138) antibody
TPT1 适用: 人 WB, ELISA 宿主: 小鼠 Monoclonal 2C4 unconjugated
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) (AA 35-138) antibodyTPT1 适用: 人 WB, ELISA 宿主: 小鼠 Monoclonal 2A3 unconjugated
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) (AA 35-138) antibodyTPT1 适用: 人 WB, ELISA, IF 宿主: 小鼠 Monoclonal 3C7 unconjugated
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) antibodyTPT1 适用: 人 WB, ELISA, IHC, IP, IF 宿主: 兔 Monoclonal 10A9 unconjugated Recombinant Antibody
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) (Internal Region) antibodyTPT1 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) antibodyTPT1 适用: 人 WB, IHC, IP 宿主: 兔 Monoclonal unconjugated
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) (AA 85-172) antibodyTPT1 适用: 人, 小鼠 WB, ELISA, IF (cc), IF (p), IHC (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) (AA 1-172) antibodyTPT1 适用: 人 WB, IHC, IP, ICC 宿主: 兔 Polyclonal unconjugated
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) (AA 1-172) antibodyTPT1 适用: 人 ELISA, IHC, IF 宿主: 兔 Polyclonal unconjugated
anti-Tumor Protein, Translationally-Controlled 1 (TPT1) (AA 85-113) antibodyTPT1 适用: 人 WB, IHC (p), FACS 宿主: 兔 Polyclonal RB19910 unconjugated
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应用备注
- Western blot: 1 μg/mL. Immunohistochemistry on paraffin sections: 5 μg/mL. Heat treatment is necessary for paraffin embedded sections. Microwave oven, 2 x for 10 minutes each in citrate buffer, pH 6.5. For details see protocols below. Not recommended for immunoprecipitation.
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实验流程
- SDS-PAGE & Western Blotting 1) Wash the cells 3 times with PBS and suspend with 10 volumes 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 an 8 mg/mL solution. 3) Mix the sample with an 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 specific 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 suggested in the APPLICATIONS for 1 hour at room temperature. (The optimal 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 1:10,000 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 (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriate chemilumi nescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 12) Expose to X-ray film in a dark room for 10 minutes. Develop the film as usual. The conditions for exposure and development may vary. Positive controls for Western blotting U937, 293T, WR19L, PC12, Rat1 Immunohistochemical staining for paraffin-embedded sections: SAB method 1) 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 treatment Heat treatment by microwave oven: Place the slides put on staining basket in 500 mL beaker with 500 mL citrate buffer ( pH 6.5). Cover the beaker with plastic wrap, then process the slides 2 times for 10 minutes each at 500 W with microwave oven. Let the slides cool down in the beaker at room temperature for about 40 minutes. 5) Remove the slides from the citrate buffer and cover each section with 3 % H 2 O 2 for 10 minutes at room temperature to block endogenous peroxidase activity. Wash 3 times in PBS for 5 minutes each. 6) Remove the slides from PBS, wipe gently around each section and cover tissues with Protein Blocking Agent for 5 minutes to block non-specific antibody staining. Do not wash. 7) Tip off the blocking buffer, wipe gently around each section and cover tissues with the primary antibody diluted with PBS containing 1 % BSA as suggested in the APPLICATIONS. 8) Incubate the sections for 1 hour at room temperature. 9) Wash the slides 3 times in PBS for 5 minutes each. 10) Wipe gently around each section and cover tissues with Polyvalent Biotinylated antibody. Incubate for 10 minutes at room temperature. Wash as in step 9. 11) Wipe gently around each section and cover tissues with Streptavidin-Peroxidase. Incubate for 10 minutes at room temper ature. Wash as in step 9. 12) Visualize by reacting for 10-20 minutes with substrate solution containing 7.5 mg DAB, 40 μ L of 30 % H 2 O 2 in 150 mL PBS. * DAB is a suspected carcinogen and must be handled with care. Always wear gloves. 13) Wash the slides in water for 5 minutes. 14) Counter stain in hematoxylin for 1 minute, wash the slides 3 times in water for 5 minutes each, and then immerse the slides in PBS for 5 minutes. Dehydrate by immersing in Ethanol 3 times for 3 minutes each, followed by immersing in Xylene 3 times for 3 minutes each. 15) Samples are now ready for mounting. Positive control for immunohistochemistry human pancreas
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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 ( pH 7.2) containing 50 % Glycerol. Contains no 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
- Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.
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- TPT1 (Tumor Protein, Translationally-Controlled 1 (TPT1))
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别名
- tpt1,tctp
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背景
- He pathophysiological events of human allergic disease are char acterized by infiltration of inflammatory cells and by the presence of mediators such as histamine. An IgE-dependen t histamine releasing factor (HRF) produced by lymphocytes of atopic children and present in biological fluids of allergic patients has been identified and purified. HRF, also known as TCTP (Translationally Controlled Tumor Protein), Fortilin, Tumor protein-1 (Tpt-1), lens epithelial protein and p23, is as a 21 or 23 kDa protein produced in a variety of mouse and human cell lines, respectively 1). HRF binds to a unique species of IgE on the surface of mast cells and basophils and thereby causes histamine release. The histamine released in late-phase reaction (LPR) of chronic allergic diseases such as asthma results from activation of basophils, eosinophils and lymphocytes infiltrate tissues during this response 2-4) .
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UniProt
- P13693
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途径
- Response to Water Deprivation, Regulation of Cell Size, Negative Regulation of intrinsic apoptotic Signaling
抗原
-
