Product Pathways - PathScan ELISA
PathScan® Total Rb Sandwich ELISA Kit #12915
|12915S||1 Kit (96 assays)||---||In Stock||---|
|12915||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
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|Kit Includes||Volume||Solution Color|
|Rb coated Microwells||96 tests|
|Biotinylated Rb Detection Antibody||11 ml||Green|
|HRP-linked Streptavidin||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.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
PathScan® Total Rb Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Rb protein. An Rb antibody has been coated onto the microwells. After incubation with cell lysates, Rb protein is captured by the coated antibody. Following extensive washing, a biotinylated Rb Antibody is added to detect the captured Rb protein. HRP-linked Streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total Rb protein.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Total Rb Sandwich ELISA Kit recognizes endogenous levels of Rb protein as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Figure 1: Treatment of serum-starved WI-38 cells (3 days) with serum for 2 days induces Rb protein expression, as detected by the PathScan® Total Rb Sandwich ELISA Kit #12915. The absorbance readings at 450 nm are shown in the top figure while the corresponding western blot using Rb (D20) Rabbit mAb #9313 is shown in the bottom figure.
The retinoblastoma tumor suppressor protein Rb regulates cell proliferation by controlling progression through the restriction point within the G1-phase of the cell cycle (1). Rb has three functionally distinct binding domains and interacts with critical regulatory proteins including the E2F family of transcription factors, c-Abl tyrosine kinase, and proteins with a conserved LXCXE motif (2-4). Cell cycle-dependent phosphorylation by a CDK inhibits Rb target binding and allows cell cycle progression (5). Rb inactivation and subsequent cell cycle progression likely requires an initial phosphorylation by cyclin D-CDK4/6 followed by cyclin E-CDK2 phosphorylation (6). Specificity of different CDK/cyclin complexes has been observed in vitro (6-8) and cyclin D1 is required for Ser780 phosphorylation in vivo (9).
- Sherr, C.J. (1996) Science 274, 1672-7.
- Nevins, J.R. (1992) Science 258, 424-9.
- Welch, P.J. and Wang, J.Y. (1993) Cell 75, 779-90.
- Hu, Q.J. et al. (1990) EMBO J 9, 1147-55.
- Knudsen, E.S. and Wang, J.Y. (1997) Mol Cell Biol 17, 5771-83.
- Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
- Connell-Crowley, L. et al. (1997) Mol Biol Cell 8, 287-301.
- Kitagawa, M. et al. (1996) EMBO J 15, 7060-9.
- Geng, Y. et al. (2001) Proc Natl Acad Sci USA 98, 194-9.
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For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.