Elisa包被液

更新:2022-1-4 15:15:06

中文名:Elisa包被液说明书下载暂无
英文名:Coating Buffer For Elisa
描述:Elisa包被液为pH9.6的即用型工作液,用于ELISA(酶联免疫吸附实验)中酶标板的抗原或抗体的包被。使用时取适量包被缓冲液将抗原或抗体溶液稀释至相应包被的浓度,在ELISA酶标板每个反应孔中加入100ul,4℃包被过夜。
订购信息
    货号规格价格品牌
    G5408-100ml100ml55GBCBIO
    G5408-500ml500ml170GBCBIO
    G5418-500ml500ml(10X)880GBCBIO
产品介绍

    间接ELisa的一般方案
    Indirect ELISA Protocol

    1. Dilute antigen to a final concentration of 1-20 μg/ml using PBS or Coating buffer. Coat the wells of a PVC microtiter plate with the antigen by pipeting 50μl of the antigen dilution in the top wells of the plate. Dilute down the plate as required. Seal the plate and incubate overnight at 4°C or 2 h at room temperature.
    2. Wash plate 3 times with PBS.
    3. Block the remaining protein-binding sites in the coated wells by adding 200 μl blocking buffer, 5% non fat dry milk/PBS, per well. Alternative blocking reagents include BlockACE or BSA.
    4. Cover the plate with an adhesive plastic and incubate for at least 2h at room temperature or, if more convenient, overnight at 4°C.
    5. Wash the plate 3 times with PBS.
    6. Add 100 μl of diluted primary antibody to each well.
    7. Cover the plate with an adhesive plastic and incubate for 2 h at room temperature.
    8. Wash the plate 4 times with PBS.
    9. Add 100 μl of conjugated secondary antibody, diluted at the optimal concentration (according to the manufacturer) in blocking buffer immediately before use.
    10. Cover the plate with an adhesive plastic and incubate for 1-2 h at room temperature.
    11. Wash the plate 5 times with PBS.
    12. Dispense 100 μl (or 50 μl) of the substrate solution per well with a multichannel pipet or a multipipet. 
    13. After sufficient color development (if it is necessary) add 50-100μl of stop solution to the wells. 
    14. Record the absorbance at 450 nm on a plate reader within 30 minutes of stopping the reaction.

    1. 使用 PBS 或包被缓冲液将抗原稀释至终浓度 1-20 μg/ml。通过在板的顶部孔中吸取 50μl 的抗原稀释液,将抗原涂在 PVC 微量滴定板的孔中。根据需要稀释板。密封板并在 4°C 下孵育过夜或在室温下孵育 2 小时。
    2. 用 PBS 洗板 3 次。
    3. 每孔加入 200 μl 封闭缓冲液、5% 脱脂奶粉(或者BSA)/PBS,封闭包被孔中剩余的蛋白质结合位点。
    4. 用封板胶密封Elisa板并在室温下孵育至少 2 小时,或者,如果更方便,在 4°C 下孵育过夜。
    5. 用 PBS 洗板 3 次。
    6. 每孔加入 100 μl 稀释的一抗。
    7. 用封板胶密封Elisa并在室温下孵育 2 小时。
    8. 用 PBS 洗板 4 次。
    9. 使用前立即加入 100 μl 偶联二抗,在封闭缓冲液中稀释至最佳浓度(根据制造商)。
    10. 用封板胶密封Elisa并在室温下孵育 1-2 小时。
    11. 用 PBS 洗板 5 次。
    12. 用多道移液管或多管移液管在每孔中分配 100 μl(或 50 μl)底物溶液。 
    13. 充分显色后(如果需要)向孔中加入 50-100μl 终止液。 
    14. 在反应停止后 30 分钟内,在酶标仪上记录 450 nm 处的吸光度。

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