Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total Survivin Sandwich ELISA Kit #7169

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

Important Ordering Details: Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.

Kit Includes Volume Solution Color
Survivin mAb Coated Microwells 96 tests
Survivin Detection Ab 11 ml Green
Anti-rabbit IgG, HRP-linked Antibody 11 ml Red
TMB Substrate #7004 11 ml Colorless
STOP Solution #7002 11 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) #9803 15 ml Yellowish

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H Mk

Reactivity Key:  H=Human  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

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 #7074 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.

Specificity / Sensitivity

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.

ELISA - Western correlation

ELISA - Western correlation

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.

Sensitivity

Sensitivity

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.

Background

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).

  1. Reed, J.C. and Reed, S.I. (1999) Nature Cell Biol. 1, 199-200.
  2. Li, F. et al. (1998) Nature 396, 580-584.
  3. Li, F. et al. (1999) Nat. Cell Biol. 1, 461-466.
  4. O'Connor, D.S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 13103-13107.
  5. Olie, R.A. et al. (2000) Cancer Res. 60, 2805-2809.
  6. Grossman, D. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 635-640.

Application References

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Protocols

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For Research Use Only. Not For Use In Diagnostic Procedures.

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