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细胞衰老检测试剂盒




货号:8058

规格:50 tests/53 mm plate

交货周期:部分现货

运杂费: 登录后可查看运杂费

DeepBio得分:5567.2
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产品说明书


产品说明书 8058

Normal mammalian cells divide for a limited number of population doublings and eventually enter an arrested state in which the cells remain alive, but do not proliferate in response to mitogens, and assume a characteristic enlarged, flattened morphology. This process, termed senescence, is thought to be a tumor suppressive mechanism and underlying cause of aging. Senescence-associated β-galactosidase (SA- β-gal) is a widely used biochemical maker for assessing senescence in cultured cells. The ScienCell™ Cell Senescence Assay provide an easy-to-use method to detect SA- β-gal by staining cells with 5-bromo-4-chloro-3-indolyl- β- D-galactopyranoside (X-gal) at pH 6.0, a pH condition that suppress lysosomal β-galactosidase activity sufficiently to ensure that nonsenescent cells remain unstained. Kit Components Cat. No. # of vials Reagent Quantity Storage 8058a 1 Staining Solution A 1 ml 4°C 8058b 1 Staining Solution B 1 ml 4°C 8058c 1 Staining Solution C 0.2 ml 4°C 8058d 1 Staining Solution D 100 ml 4°C 8058e 1 X-gal Solution 5 ml -20°C 8058f 1 100× Fixing Solution 1 ml 4°C Quality Control The Cell Senescence Assays are applied to early passage and senescent ScienCell™ Human Renal Proximal Tubular Epithelial Cells (HRPTEpiCs). Data show that the senescent cells show positive SA- β-gal staining while most early passage cells don’t (Figure 1). Procedures A. Preparation of reagents 1. Preparation of working fixing solution: Prepare 1× fixing solution by diluting 100X Fixing Solution stock 1:100 in PBS. 2. Preparation of working staining solution: Prepare fresh staining working solution based on the number of samples to be assessed. For each sample in 35 mm plate, prepare the following mixture: 100 µl of X-gal Solution 20 µl of Staining Solution A 20 µl of Staining Solution B 4 µl of Staining Solution C + 1856 µl of Staining Solution D 2000 µl of working staining solution B. Staining protocol 2 1. Remove culture medium from cells and rinse twice with PBS. 2. Fix cells by incubating with 2 ml of working fixing solution for 3-5 minutes at room temperature. 3. Aspirate working fixing solution and rinse the fixed cells three times with PBS. 4. Add 2 ml of working staining solution to completely cover cells and incubate cells at 37°C, protected from light, for 12-24 hours, blue color should develop in senescent cells.* Examine cells at regular time points to avoid overstraining. 5. After incubation, remove working staining solution and rinse cells twice with PBS, keep the cells in PBS at 4°C. Examine and count the blue stained cells using a light microscope. * Crystal deposition, which comes from unreacted X-gal, may be observed after incubation of cells with working staining solution. It can be minimized by pre-filtering the working staining solution with a 0.2 µm filter. Figure 1. Human renal proximal tubular epithelial cells (HRPTEpiCs) probed with ScienCell™ Senescence Assay. Most senescent cells show positive SA-β-Gal staining (A), while only a few labeled cells can be observed in early passage culture (B).