HeLa Ser/Thr Phosphatase-Inhibited Whole Cell Lysate Set: Calyculin-Treated and Vehicle-Treated (ab135395)

Overview

  • Product name
    HeLa Ser/Thr Phosphatase-Inhibited Whole Cell Lysate Set: Calyculin-Treated and Vehicle-Treated
  • Product overview

    HeLa whole cell extracts were collected in lysis buffer after 3 hour treatment with Calyculin A (50 nM) or vehicle (DMSO) in serum-free media. The protein content was determined by BCA assay. Calyculin A is a potent phosphatase inhibitor. Unlike Okadaic Acid, which only acts on protein phosphatase PP2A, Calyculin A inhibits both PP1 and PP2A. Cells treated with Calyculin A accumulate serine/threonine phosphorylation making them a strong positive control for phospho-threonine or phospho-serine specific antibodies.

     

    Concentration:

    HeLa  Lysate - Vehicle Treated Control - 200 µg at 1 mg/mL

    HeLa  Lysate - Calyculin Treated - 200 µg at 1 mg/mL

     

    The lysate is supplied in SDS-PAGE loading buffer with DTT and is ready to load on a gel.

  • Tested applications
    Suitable for: WB, SDS-PAGEmore details

Properties

    Applications

    Our Abpromise guarantee covers the use of ab135395 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    WB Use at an assay dependent concentration.
    SDS-PAGE Use at an assay dependent concentration.

    HeLa Ser/Thr Phosphatase-Inhibited Whole Cell Lysate Set: Calyculin-Treated and Vehicle-Treated images

    • All lanes: Anti-Phosphoserine/Threonine antibody (ab17464) at 1/1000

      Lane 1: Marker
      Lane 2: HeLa Whole Cell Lysate - Calyculin Treated at 20 µg
      Lane 3: HeLa Whole Cell Lysate – Untreated Control at 20 µg


      Secondary:
      Goat polyclonal to Rabbit IgG – Pre-Adsorbed (HRP) at 1/6000 developed using the ECL technique.

      Performed under reducing conditions.

    • Lane 1: Marker
      Lane 2: HeLa Whole Cell Lysate - Calyculin Treated at 20 µg
      Lane 3: HeLa Whole Cell Lysate – Untreated control at 20 µg


      Gel
      10-20% acrylamide separation gel

    References for HeLa Ser/Thr Phosphatase-Inhibited Whole Cell Lysate Set: Calyculin-Treated and Vehicle-Treated (ab135395)

    ab135395 has not yet been referenced specifically in any publications.

    Product Wall



    1. What is the lysis buffer that is used to make this extract? Is it a hypotonic lysis buffer, or do you use detergents (like Triton or Tween)?


    - I can confirm that for ab135395 the cells are directly lysated in SDS buffer. Sinc...

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