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- 详细信息
- 文献和实验
- 技术资料
- 库存：
大量
- 英文名：
Anti-alpha-Synuclein (Rat) ELISA Kit *Colorimetric*
- CAS号：
Anti-alpha-Synuclein (Rat) ELISA Kit *Colorimetric*
- 保质期：
详见说明
- 供应商：
齐源生物
- 保存条件：
4℃
- 规格：
1 kit
α-Synuclein Quantitative ELISA Kit (Rat) 提供了一种方便的定量测定方法，用于确定细胞和组织裂解物以及体液中的 α-Synuclein 含量。与市场上的其他抗 α-突触核蛋白 ELISA 试剂盒相比，运行该检测所需的时间更短。本试剂盒中的 HRP 偶联检测抗体在检测过程中与样品和标准品同时添加。这消除了额外的孵育和洗涤步骤，并使该试剂盒成为 α-突触核蛋白定量的一步程序。检测限：5 pg/ml、主持人：大肠杆菌、来源/物种：大鼠
1 Reconstitute rat α-Synuclein Standard (Component B) with 1 ml of Sample Dilution Buffer
(Component C) or Sample Dilution Buffer mixed with lysis solution (see Note “d”). Mix gently (do not
vortex) and let stand for 10-15 minutes. Reconstituted standard must be used within one hour to
ensure accurate results. Do not re-use the reconstituted Standard!
2 Arrange and label strips (Component A) based on the number of wells for standard and samples.
Although diluted standard and samples can be run as single points, duplicates are recommended.
Instructions for preparing cell and/or tissue lysates are provided in the Appendix.
Place unused strips into the plate bag and seal completely.
3 Make serial dilution of rat α-Synuclein Standard (Component B) with Sample Dilution Buffer
(Component C). Refer to Table 1.
4 Dilute Detection Antibody (Component G) 200 fold in Sample Dilution Buffer (Component C). Prepare
50 μl of the above for each well to be run in the assay.
5 Add 100 μl of the diluted standards in duplicates including blank (Start with Step 3 from Table 1). We
recommend to dilute rat plasma at 1:50 ratio with Sample Dilution Buffer to avoid
sample matrix effect. In addition, protease inhibitor AEBSF should be added to all samples at 1mM
final concentration to avoid protein degradation.
6 Add 100 μl per well of the diluted samples into appropriate wells (depending on the number of
samples to be tested).
7 Add 50 μl of the diluted Detection Antibodies (from step 1.4) into each well to be tested, cover the
plate with Adhesive Plate Cover (Component H), and incubate it at room temperature for 4 hours or
4 C overnight. Protect plate from direct light!
1.8 Prepare 1X working wash buffer by diluting the 10X Wash Buffer (Component D) with deionized H2O.
9 After plate incubation, aspirate the wells and wash them with 350 μl/well of 1x Wash Buffer 6 times.
Allow 5-10 seconds lag time before emptying the wells between washes. Pat dry the plate using a
paper towel and clean outside of wells with non-abrasive paper to ensure accurate optical reading.
10 Add 100 μl of the TMB color substrate solution (Component E) into each well. Incubate plate
at room temperature until blue gradient is clearly observed across the wells (5-15 minutes). It may
be necessary to adjust color development time so that absorbance values fall within the detection
range. Samples may require further dilution if α-Synuclein concentration is too high.
11 Add 100 μl of the Stop Solution (Component F) into each well (blue color will turn to yellow). Measure
absorbance (OD) at 450 nm using a microplate absorbance reader within 20 minutes after adding
the Stop Solution.
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文献和实验人α-突触核蛋白(a-Synuclein)ELISA试剂盒 说明书
上海西唐生物科技有限公司 021-55229872, 65333639 www.westang.com 人 α- 突触核蛋白 ( a -Synuclein)ELISA 试剂盒 ( 用于血清、血浆、细胞培养上清液和其它生物体液内 ) 原理 本实验采用双抗体夹心 ABC-ELISA 法。用抗人 a -Synuclein 单抗包被于酶标板上，标准品和样品中的
大鼠海马体神经元和活体大鼠脑注射 α 突触核蛋白后做的免疫细胞化学和免疫组化分析。 原代大鼠海马体神经元被 1 型突触核蛋白前体原纤维（SPR-322）4 µg/mL（D-F）处理后显示路易体内含物的形成，而当被 2 型突触核蛋白前体原纤维（SPR-317）4 µg/mL（A-C）处理时没有路易体内含物形成。组织：原代海马体神经元，物种：斯普拉-道来氏大鼠，固定：由 PFA 制作的 4% 甲醛。一抗：小鼠抗 pSer129 抗体, 稀释度 1:1,000，4 °C 下处理 24 小时；二抗
用于免疫检测的特殊配方的抗体稀释液，可以促进抗原抗体特异性结合，产生比传统方法高几倍或十几倍的信号强度，从而解决免疫检测实验中（比如免疫印迹，ELISA等）经常遇到的低灵敏度和高背景强度等问题，得到准确清晰的实验结果。 试剂盒操作简单方便，含有溶液1和2两种即用型溶液，分别用做一抗和二抗的稀释液，替代其他的传统稀释缓冲液，包括TBS，TBST，PBS，PBST等。其余操作均按照标准的免疫检测操作流程进行。此外，试剂盒中各组分对标记抗体的酶活性（辣根过氧化物酶HRP，碱性磷酸酶AP等）均没有影响
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