Figure 1: Phospho-p90RSK (Thr359) protein from untreated (-) and TPA treated (+) HeLa cells is detected by PathScan® Phospho-p90RSK (Thr359) Sandwich ELISA kit. The absorbance readings at 450 nm are shown in the top figure,
while the corresponding Western blot using Phospho-p90RSK (Thr359) (D1E9) Rabbit mAb #8753 is shown in the bottom figure.
Figure 2. The relationship between the protein concentration of lysates from untreated or TPA-treated HeLa cells, and the absorbance at 450 nm as detected by PathScan® Phospho-p90RSK (Thr359) Sandwich ELISA Kit is shown. Starved HeLa cells (85% confluent) were untreated or treated with TPA #4174 (200 nM, 15 minutes, 37ºC), and then lysed with cell lysis buffer.
|REACTIVITY||H R Mk|
|Product Includes||Volume||Solution Color|
|Phospho-p90RSK (Thr359) Rabbit mAb Coated Microwells||96 tests|
|RSK Mouse Detection mAb||1 ea||Green (Lyophilized)|
|Anti-mouse 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|
The PathScan® Phospho-p90RSK (Thr359) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of p90RSK protein phosphorylated at Thr359. A phospho-p90RSK (Thr359) rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-p90RSK proteins can be captured by the coated antibody. Following extensive washing, a p90RSK mouse antibody is added to detect the captured phospho-p90RSK protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of p90RSK phosphorylated at Thr359.
Antibodies in this 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
PathScan® Phospho-p90RSK (Thr359) Sandwich ELISA Kit detects endogenous levels of p90RSK protein phoshorylated at Thr359 in human cells, 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.
Human, Rat, Monkey
The 90 kDa ribosomal S6 kinases (RSK1-4) are a family of widely expressed Ser/Thr kinases characterized by two nonidentical, functional kinase domains (1) and a carboxy-terminal docking site for extracellular signal-regulated kinases (ERKs) (2). Several sites both within and outside of the RSK kinase domain, including Ser380, Thr359, Ser363, and Thr573, are important for kinase activation (3). RSK1-3 are activated via coordinated phosphorylation by MAPKs, autophosphorylation, and phosphoinositide-3-OH kinase (PI3K) in response to many growth factors, polypeptide hormones, and neurotransmitters (3).
Explore pathways + proteins related to this product.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.