Overview

  • Product name

    Calyculin A, protein phosphatase inhibitor
  • Description

    Potent, selective and cell-permeable protein phosphatase inhibitor
  • Biological description

    Potent, selective and cell-permeable protein phosphatase inhibitor (IC50 values are 2 and 1 nM for PP1 and PP2A, respectively). Highly selective over PP2B and PP2C. Active in vivo and in vitro.
  • Purity

    > 99%
  • CAS Number

    101932-71-2
  • Chemical structure

    Chemical Structure

Properties

  • Chemical name

    [(2R,3R,5R,7R,8S,9S)-2-[(1S,3S,4S,5R,6R,7E,9E,11E,13Z)-14-Cyano-3,5-dihydroxy-1-methoxy-4,6,8,9,13-pentamethyltetradeca-7,9,11,13-tetraenyl]-9-[(E)-3-[2-[(2S)-4-[[(2S,3S,4S)-4-(dimethylamino)-2,3-dihydroxy-5-methoxypentanoyl]amino]butan-2-yl]-1,3-oxazol-4-yl]prop-2-enyl]-7-hydroxy-4,4,8-trimethyl-1,10-dioxaspiro[4.5]decan-3-yl] dihydrogen phosphate
  • Molecular weight

    1009.18
  • Molecular formula

    C50H81N4O15P
  • PubChem identifier

    5311365
  • Storage instructions

    Store at -20°C. Store under desiccating conditions. The product can be stored for up to 12 months.
  • Solubility overview

    Soluble in ethanol and in DMSO
  • Handling

    Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one month. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.

    Toxic, refer to SDS for further information.

    Need more advice on solubility, usage and handling? Please visit our frequently asked questions (FAQ) page for more details.

  • SMILES

    CC1C(CC2(C(C(C(O2)C(CC(C(C)C(C(C)C=C(C)C(=CC=CC(=CC#N)C)C)O)O)OC)OP(=O)(O)O)(C)C)OC1CC=CC3=COC(=N3)C(C)CCNC(=O)C(C(C(COC)N(C)C)O)O)O
  • Source

    Discodermia calyx

  • Research areas

Images

  • Calyculin A inhibits the growth of breast cancer epithelial MCF7 cells. Cells were incubated with different concentrations of Calyculin A (ab141784) for four days. Cell number was measured using the methylene blue method. The number of cells was normalized with respect to the control (100%) and plotted against Calyculin A concentrations.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling NF-kB p65 (phospho S276) with ab183559 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    The expression increased on HeLa cells after treatment with Calyculin A (ab141784 100ng/ml, 10min) then TNF-a (20ng/ml, 5min).

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • All lanes : Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) grown in serum-free media overnight, whole cell lysate
    Lane 2 : HeLa grown in serum-free media overnight, then treated with 100 ng/ml Calyculin A (ab141784) for 15 minutes, followed by Calyculin A removal and treatment with 100 ng/ml BMP2 for 30 minutes, whole cell lysate
    Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) grown in serum-free media overnight, whole cell lysate
    Lane 4 : NIH/3T3 cultured in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Observed band size: 60 kDa
    why is the actual band size different from the predicted?



    Exposure time:
    Lanes 1 and 2: 3 minutes.
    Lanes 3 and 4: 30 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • NF-kB p65 (phospho S276) was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates, treated with 100ng/ml Calyculin A (ab141784) for 10min, then 20ng/ml TNA-a for 5min, with ab183559 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab183559 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate treated with 100ng/ml Calyculin A (ab141784) for 10min, then 20ng/ml TNA-a for 5min, 10 μg (Input).

    Lane 2: ab183559 IP in HeLa whole cell lysate treated with 100ng/ml Calyculin A (ab141784) for 10min, then 20ng/ml TNA-a for 5min.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab183559 in HeLa whole cell lysate treated with 100ng/ml Calyculin A (ab141784) for 10min, then 20ng/ml TNA-a for 5min.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

  • All lanes : Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559) at 1/1000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa whole cell lysate treated with 100ng/ml Calyculin A ab141784 for 30 minutes, then treated with 20ng/ml TNF-a for 5 minutes
    Lane 3 : HeLa whole cell lysate treated with 100ng/ml Calyculin A ab141784 for 30 minutes, then treated with 20ng/ml TNF-a for 5 minutes, then treated with Alkaline Phosphatase for 1 hour

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
  • All lanes : Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) grown in serum-free media overnight, whole cell lysate
    Lane 2 : HeLa grown in serum-free media overnight, then treated with 100ng/ml Calyculin A (ab141784) for 15 minutes, Calyculin A was removed, followed by treatment with 100ng/ml BMP2 for 30 minutes, whole cell lysate
    Lane 3 : NIH/3T3 (mouse embryonic fibroblast) grown in serum-free media overnight, whole cell lysate
    Lane 4 : NIH/3T3 grown in serum-free media overnight, then treated with 50ng/ml BMP2 for 30 minutes, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 60 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution: 5% NFDM/TBST.

  • All lanes : Anti-AS160 (phospho T642) antibody [EPR2733(2)] (ab131214) at 1.12 µg/ml

    Lane 1 : HEK-293 (human embryonic kidney epithelial cell) grown in serum free media overnight whole cell lysate
    Lane 2 : HEK-293 grown in serum free media overnight, then treated with 100nM Calyculin A (ab141784) for 50min and then 100ng/ml Insulin was added for the last 20min, whole cell lysate
    Lane 3 : HEK-293 grown in serum free media overnight, then treated with 100nM Calyculin A (ab141784) for 50min and then 100ng/ml Insulin was added for the last 20min, whole cell lysate. Then the membrane was incubated with alkaline phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution


    Blocking and diluting buffer: 5% NFDM/TBST. 

References

This product has been referenced in:

  • Bruno MK  et al. Inhibition of protein phosphatase activity and changes in protein phosphorylation following acetaminophen exposure in cultured mouse hepatocytes. Toxicol Appl Pharmacol 153:119-32 (1998). Read more (PubMed: 9875306) »
  • Ishihara H  et al. Calyculin A and okadaic acid: inhibitors of protein phosphatase activity. Biochem Biophys Res Commun 159:871-7 (1989). Read more (PubMed: 2539153) »
  • Ishihara H  et al. Calcium-independent activation of contractile apparatus in smooth muscle by calyculin-A. J Pharmacol Exp Ther 250:388-96 (1989). Read more (PubMed: 2545866) »
See all 2 Publications for this product

Customer reviews and Q&As

Answer



Ich habe nun mehr Informationen vom Labor und kopiere diese direkt hier in die E-Mail für Sie:

"The Hela cell line was cultured in the size of 150cm²cell culture flask (#430825) from corning company. The cells were uniformly distributed in the 150cm²cell culture flask. The Hela cells were serum-starved for overnight and then the Hela cells were treated with Calyculin A 100ng/ml (work concentration) for 30min at 37C.

It’s about 5ul Calyculin A (from the stock solution at 100ug/ml) added into the 5ml cell supernatant/ medium in the flask. The cells were harvested after treatment."

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