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.

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

ab135395 has not yet been referenced specifically in any publications.

Customer reviews and Q&As



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...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up