Figure 1: Survivin protein from human cells (HeLa, H526 and MCF-7) and monkey cells (COS) can be detected by PathScan® Total Survivin Sandwich ELISA Kit #7169. However, this kit cannot detect the survivin protein from other species, such as mink (MV1LU), rat (C6), mouse (BAF3) and hamster (CHO) cell lines. The absorbance readings at 450 nm are shown in the top figure, while the corresponding Western blot using Survivin (6E4) Mouse mAb #2802, is shown in the bottom figure.
Figure 2: The relationship between protein concentration of lysates from HeLa cells and the absorbance at 450 nm is shown. HeLa cells (80% confluence) were harvested and then lysed.
|Product Includes||Volume||Solution Color|
|Survivin Mouse mAb Coated Microwells||96 tests|
|Survivin Rabbit Detection mAb||1 ea||Green (Lyophilized)|
|Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|TMB Substrate 7004||11 ml|
|STOP Solution 7002||11 ml|
|Sealing Tape||2 ea|
|ELISA Wash Buffer (20X) 9801||25 ml|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) 9803||15 ml|
CST's PathScan® Total Survivin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total survivin protein. A Survivin Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho and nonphospho survivin proteins are captured by the coated antibody. Following extensive washing, Survivin Rabbit Detection Antibody is added to detect the captured survivin protein. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of total survivin protein.
Antibodies in kit are custom formulations specific to kit.
NOTE: Prepare solutions with purified water.
*NOTE: Some PathScan® ELISA Kits may include HRP-Linked Streptavidin in place of HRP-Linked Antibody.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
posted November 2013
Protocol Id: 204
CST's PathScan® Total Survivin Sandwich ELISA Kit #7169 detects endogenous levels of total survivin protein. As shown in Figure 1, survivin protein from human (HeLa, H526 and MCF-7) and monkey (COS), is detected by this kit. In Figure 3, Western analysis of protein captured in microwells coated with the survivin capture antibody shows a major band corresponding to the survivin protein (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Survivin is a 16 kDa anti-apoptotic protein highly expressed during fetal development and cancer cell malignancy (1). Survivin binds and inhibits caspase-3, controlling the checkpoint in the G2/M-phase of the cell cycle by inhibiting apoptosis and promoting cell division (2,3). This regulatory process requires the phosphorylation of survivin at Thr34 by p34 cdc2 kinase (4). Gene targeting using a Thr34 phosphorylation-defective survivin mutant, as well as antisense survivin, have been shown to inhibit tumor growth (5,6).
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