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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.
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.
|HeLa Whole Cell Lysate - Calyculin Treated||1 x 200µg|
|HeLa Whole Cell Lysate - Vehicle Treated Control||1 x 200µg|
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.
|WB||Use at an assay dependent concentration.|
|SDS-PAGE||Use at an assay dependent concentration.|
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
Goat polyclonal to Rabbit IgG – Pre-Adsorbed (HRP) at 1/6000 developed using the ECL technique.
Performed under reducing conditions.
ab135395 has not yet been referenced specifically in any publications.