Interested in promotions? | Click here >>
7136
PathScan® Cell Growth 4-Plex Array Kit
ELISA Kits
Antibody Array Kit

PathScan® Cell Growth 4-Plex Array Kit #7136

Citations (0)
PathScan® Cell Growth 4-Plex Array Kit: Image 1

Figure 1: Schematic representation of the position of targets in a single well.

PathScan® Cell Growth 4-Plex Array Kit: Image 2

Figure 2: NIH/3T3 cells were starved and stimulated with PDGF (100 ng/ml) for 5 or 40 minutes. Lysates were analyzed using the PathScan® Cell Growth 4-Plex Array Kit. Plate analysis was performed using a Quansys Biosciences Imager. Spot intensity was quantified using Q-View™ software. The bar graph shows the measurements of four different target proteins. Corresponding Western blots obtained with the same cell lysates are shown at the bottom.

PathScan® Cell Growth 4-Plex Array Kit: Image 3

Figure 3: Cell extracts from unstimulated or stimulated NIH/3T3 cells were applied to wells at increasing total protein concentrations and analyzed. Luminescent signals were acquired and spot intensity quantified. The graphs show the relationship between the total protein concentration and the resulting strength of the luminescent signal for each target.

To Purchase # 7136

Product Description

CST’s PathScan® Cell Growth 4-Plex Array Kit is based upon the sandwich ELISA principle. Rather than one immobilized antibody, these PathScan® 4-Plex Array Kits contain four different antibodies in each well of a 96-well plate. This allows the researcher to measure four different target proteins from a single well simultaneously. In addition, the incorporation of multiple phospho-specific antibody pairs allows the researcher to measure the activation state of important signaling nodes. The PathScan® Cell Growth 4-Plex Array Kit allows the detection of: Phospho-Akt1 (Ser473); Phospho-S6 Ribosomal Protein (Ser235/236); Phospho-p44 MAPK (Thr202/Tyr 204); and Total Akt1 protein.

Briefly, 4 capture antibodies with distinct target specificity have been spotted onto the bottom of each well of a microwell plate. After incubation with cell lysates, the spotted antibodies capture the target proteins. Following extensive washing, a mixture of detection antibodies is added to detect the captured target proteins. An HRP-linked secondary solution is then used to recognize the bound detection antibodies. Chemiluminescent HRP substrate is used to produce luminescent signal.

Storage

Kit should be stored at 4°C with the exception of Lysis Buffer, which is stored at –20°C (packaged separately).

Specificity / Sensitivity

PathScan® Cell Growth 4-Plex Array Kit detects endogenous levels of all target proteins. As shown in Figure 2, stimulation of NIH/3T3 cells with PDGF promotes phosphorylation of Akt1 at Ser473, S6 ribosomal protein at Ser235/236 and p44 MAP kinase at Thr202/Tyr204. Levels of total Akt1 remain unchanged.

Background

Cells must respond in an appropriate fashion to many complex signaling events. Extracellular signaling cues are organized into well defined signal transduction modules that control fundamental cellular behavior. Two prominent signaling modules that are among the best characterized are the p44/42 MAP kinase (ERK MAPK) and Akt signal transduction pathways. These signaling modules control cellular growth, proliferation, movement, survival and death. The p44/42 MAPK is activated by a wide variety of extracellular signals including growth and neurotrophic factors, cytokines, hormones and neurotransmitters. p44/42 MAPK activation occurs through phosphorylation of threonine and tyrosine at the sequence T*EY* by a dual specificity kinase called MAP kinase kinase (MEK). The Akt protein kinase is generally activated in response to growth-factor stimulation and phosphorylated on Ser473. Growth factor stimulation results in phosphorylation of S6 ribosomal protein on Ser235/236, leading to an increase in protein synthesis and cell cycle progression. Phosphorylation levels of critical molecular switches such as MAPs and Akt therefore serve as a reliable indicator of the activation state of the entire signaling module. The profiling of phosphorylation events using phospho-specific antibodies is now widely used to investigate diagnostic pathology (1,2). For example, increased levels of Akt phosphorylated at Ser473 coupled with decreased levels of phospho-p44/42 predicts poor clinical outcome in prostate cancer patients (3). The PathScan® Cell Growth 4-Plex Array Kit provides the researcher with means to profile numerous chemical compounds and obtain in-cell relative potency (4).

  1. Mandell, J.W. (2003) Am J Pathol 163, 1687-98.
  2. Sheehan, K.M. et al. (2008) Oncogene 27, 323-31.
  3. Kreisberg, J.I. et al. (2004) Cancer Res 64, 5232-6.
  4. Gechtman Z. (2006) American Drug Discovery 1:1, 44-52.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.