细胞与组织裂解液(一氧化氮检测用)

C752147-100ml

100ml

阿拉丁的细胞与组织裂解液(一氧化氮检测用)(Cell and Tissue Lysis Buffer for Nitric Oxide Assay)是一种专门用于一氧化氮或总一氧化氮检测的细胞与组织裂解液。使用本裂解液裂解的样品，可以用于阿拉丁生产的一氧化氮检测试剂盒、总一氧化氮检测试剂盒的检测。本裂解液裂解获得的样品中包含了细胞或组织样品中的大部分蛋白，可以用阿拉丁生产的BCA蛋白浓度测定试剂盒测定蛋白浓度，并可以用于SDS-PAGE和Western印迹检测。需要注意的是本裂解液中不含蛋白酶和磷酸酯酶的抑制剂。本裂解液中添加蛋白酶和磷酸酯酶抑制剂可能会干扰后续的一氧化氮检测。本裂解液可以耐受反复冻融。本裂解液如果用于培养于六孔板中一个孔的细胞的裂解，或者用于20mg组织的裂解液，约可以裂解500-1000个样品。 注意事项 ： 裂解样品的所有步骤都需在冰上或4℃进行。本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。 使用说明 ： 1.对于培养的细胞样品： a.融解细胞与组织裂解液(一氧化氮检测用)，混匀备用。 b. 对于贴壁细胞：去除培养液，用PBS、生理盐水或无血清培养液洗一遍。按照6孔板每孔加入100-200微升裂解液的比例加入裂解液。用枪吹打数下，使裂解液和细胞充分接触。通常裂解液接触细胞1-2秒后，细胞就会被裂解。 c. 对于悬浮细胞：离心收集细胞，用手指把细胞用力弹散。按照6孔板每孔细胞加入100-200微升裂解液的比例加入裂解液。再用手指轻弹以充分裂解细胞。充分裂解后应没有明显的细胞沉淀。如果细胞量较多，必需分装成50-100万细胞/管，然后再裂解。大团的细胞较难裂解充分，而少量的细胞由于裂解液容易和细胞充分接触，相对比较容易裂解充分。 d.充分裂解后，10000-14000g离心3-5分钟，取上清，即可进行后续的一氧化氮检或蛋白浓度测定等操作。样品制备后如果当日不能完成一氧化氮检测，可以-20℃保存，但仍宜尽快完成检测。 裂解液用量说明：通常6孔板每孔细胞加入100微升裂解液已经足够，但如果细胞密度非常高可以适当加大裂解液的用量到150微升或200微升。2.对于组织样品： a.融解细胞与组织裂解液(一氧化氮检测用)，混匀备用。 b.把组织剪切成细小的碎片。 c.按照每20毫克组织加入100-200微升裂解液的比例加入裂解液。(如果裂解不充分可以适当添加更多的裂解液，如果需要高浓度的样品，可以适当减少裂解液的用量。) d.用玻璃匀浆器或其它适当的匀浆设备匀浆，直至充分裂解。(如果组织样品本身非常细小，加入裂解液后可以直接通过强烈vortex使样品裂解充分。直接裂解的优点是比较方便，不必使用匀浆设备，缺点是不如使用匀浆设备那样裂解得比较充分。) e.充分裂解后，10000-14000g离心3-5分钟，取上清，即可进行后续的一氧化氮检或蛋白浓度测定等操作。样品制备后如果当日不能完成一氧化氮检测，可以-20℃保存，但仍宜尽快完成检测。 Precautions ： All procedures should be performed on ice or at 4℃This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation. Instructions for Use ： 1. Preparation of cell samples a. Thaw Cell and Tissue Lysis Buffer for Nitric Oxide Assay and mix well before use.b. For adherent cells: Remove the culture medium and wash the cells once with PBS, saline, or serum-free medium. Add 100-200μl of this product to each well of a 6-well plate and pipette several times to ensure sufficient contact of cells with the lysis buffer. Cells are typically lysed after 1-2 seconds.c. For suspension cells: Collect cells by centrifugation, vortex gently or flick the bottom of the tube to disperse cells. Add 100-200μl of this product to each well of cells in 6-well plates, flick the bottom of the tube to resuspend and lyse cells thoroughly. For large amounts of cells, dispense them into 0.5-1.0×106 cells per tube for lysis.d. After complete lysis, centrifuge at 10,000-14,000×g for 3-5 minutes, and take the supernatant for NO assay or protein concentration determination. If the NO assay cannot be completed on the same day after sample preparation, store the supernatant at -20℃ and complete the assay as soon as possible.Note: Usually, 100μl of this product for each well of cells in 6-well plates is sufficient, but if the cell density is very high, the amount of lysis buffer can be increased to 150μl or 200μl.2. Preparation of tissue samples a. Thaw Cell and Tissue Lysis Buffer for Nitric Oxide Assay and mix well before use.b. Cut the tissue into small pieces.c. Add 100-200μl of this product per 20mg of tissue. The amount of lysis buffer can be adjusted based on the lysis results.d. Homogenize tissues thoroughly with the TissueMasterTM Handheld Homogenizer (, E6600) or other suitable homogenization devices. For tiny tissues, add lysis buffer after appropriate cutting and lyse with vigorous vortex. Centrifuge and take the supernatant for subsequent analysis. This lysis method is convenient, with no homogenization needed, but the lysis result is not as thorough as that from the homogenization method.e. After complete lysis, centrifuge at 10,000-14,000×g for 3-5 minutes, and take the supernatant for NO assay or protein concentration determination. If the NO assay cannot be completed on the same day after sample preparation, store the supernatant at -20℃ and complete the assay as soon as possible.