BioPath Online

Pathway Focus: The Akt Pathway

Cell Staining Simulation tool

Immunoassays Selection Guide

3D Cell and Cell Staining Simulation tool iPad app

Quickly Screen Markers In the Akt Pathway—New Magnetic Akt 7-plex Panels

  • Save sample and time―measure seven targets simultaneously in one well
  • Superior performance―accurate, reproducible, and sensitive quantitation of multiple proteins
  • Precision―high reproducibility and consistency (<10% CV)


The new Akt Pathway Phospho and Total 7-Plex Magnetic Panels provide accurate quantitation of seven proteins in cell lysates and tissue homogenates. These panels are suitable for use with the Luminex® 100/200™, FLEXMAP 3D®, and MAGPIX® systems.

Akt is one of the most actively studied kinases in both the basic research and drug development arenas. Activated Akt plays a key role in mediating signals for cell growth, cell survival (anti-apoptotic), cell cycle progression, differentiation, transcription, translation, and glucose metabolism.

Magnetic bead–based Luminex® assays simplify assay wash steps and maximize uniformity of results. They also enable automation, decrease hands-on time, and increase throughput and precision. Easy-to-use protocols enable users to obtain results in as little as 3.5 hours.

 

 Figure 1. To evaluate the performance of this kit, MCF-7 cells, insulin receptor (IR)–transfected CHO-T cells, and Jurkat cells were treated with insulin or lithium chloride (LiCl) to stimulate the phosphorylation of the Akt pathway analytes.

For Research Use Only. CAUTION: Not intended for human or animal therapeutic or diagnostic use.

New GSK3beta ABfinity™ Recombinant Antibody

  • ABfinity™ rabbit recombinant antibodies are screened to be sensitive and specific
  • Enables consistent performance for your assays
  • Consistent product from lot to lot


ABfinity™ recombinant rabbit antibodies are produced from specific recombinant clones to enable consistent antibody performance over time. ABfinity™ antibodies are available against many targets, including AKT pathway proteins.

GSK3beta belongs to the subfamily of glycogen synthase kinases, which play a central role in energy metabolism, neuronal cell development, and body pattern formation. GSK3alpha and GSK3beta are two isoforms (51 kDa and 47 kDa, respectively) that are not functionally identical. GSK3beta plays a key role in insulin and Wnt signaling. In response to insulin, PI3K and Akt/PKB are activated, and in turn inhibit GSK3beta by enhancing phosphorylation at serine 9. The result is decreased phosphorylation of GSK3beta substrates, including glycogen synthase eIF-2B, and increased glycogen synthesis and protein synthesis. ABfinity™ recombinant monoclonal antibodies, including ABfinity™ GSK3beta antibody, are ideal reagents for signaling pathway research. The ABfinity™ technology helps ensure consistent antibody performance so you don’t have to re-optimize your assay with each new lot.

We offer an extensive antibody portfolio to study the AKT signaling pathway:

 

 Figure 2. Western blot analysis was performed on whole cell extracts: HeLa (lane 1), HEK293 (lane 2), HepG2 (lane 3), Jurkat (lane 4), MCF7 (lane 5), Colo-205 (lane 6), A431 (lane 7), Cos7 (lane 8), and DU-145 (lane 9). Endogenous GSK3beta at ~47 kDa was detected using GSK3beta (GSK3B) Rabbit mAb (1H9L3) ABfinity™ Recombinant Monoclonal Antibody at 0.1 μg/mL. The blot was developed using a chemiluminescence (ECL) method.

For Research Use Only. CAUTION: Not intended for human or animal therapeutic or diagnostic use.

A Common Fix for Your Immunofluorescence Samples—Image-iT® Fixation/Permeabilization Kit

  • A complete fixation/permeabilization kit with ready-to-use components and room temperature storage
  • Optimized for secondary antibody labeling, fixed-cell dye staining, and GFP applications
  • Compatible with analysis of most cellular antigens


The Image-iT® Fixation/Permeabilization Kit contains all the necessary reagents to prepare your cells for immunocytochemical or fixed-cell dye staining applications. To streamline the workflow, we have packaged each of the components at 1X concentration in a neatly designed box with an easy-to-follow protocol printed on the inside flap so it’s always handy when you need it. And, the Image-iT® Fixation/Permeabilization Kit is stable at room temperature for 18 months. This means that you no longer have to order multiple reagents and then dilute, aliquot, and store them separately under conditions that are not necessarily optimal.

Cells are treated sequentially with the fixative, permeabilization solution, and blocking buffer according to a simple protocol. The reagents have been rigorously tested for compatibility with both nuclear and cytoplasmic targets and deliver results as good as or better than standard reagents. Care was taken to optimize the kit for preservation of 3D cellular structure and adequate antibody access to intracellular antigenic sites. The high-quality components and optimized formulations help decrease background staining, consequently improving signal-to-noise levels and image quality.

 

Figure 3. (A) Human dermal fibroblasts, neonatal (HDFn) (C-004-5C), were fixed and permeabilized using the Image-iT® Fixation/Permeabilization Kit. Mitochondria were labeled with an anti-CVIP antibody and stained with an Alexa Fluor® 488 goat anti-mouse secondary antibody. Actin was stained using Alexa Fluor® 594 phalloidin, and nuclei using Hoechst 33342. Images were acquired on a Nikon upright microscope (60x). (B) BPAE cells were fixed and permeabilized using the Image-iT® Fixation/Permabilization Kit, then labeled with an anti–beta-tubulin antibody and stained with an Alexa Fluor® 488 goat anti-mouse secondary antibody and Click-iT® EdU Alexa Fluor® 647. Images were acquired on a Nikon upright microscope (60x).

For Research Use Only. CAUTION: Not intended for human or animal therapeutic or diagnostic use.

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The trademarks mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. Luminex, FLEXMAP 3D, and MAGPIX are registered trademarks of Luminex Corporation.