Overview

  • Product name
    Staurosporine, Protein kinase inhibitor
  • Description
    Protein kinase inhibitor
  • Biological description
    Potent, cell-permeable, broad spectrum inhibitor of protein kinases. Kinases inhibited include: protein kinase C (IC50 = 3 nM), cAMP-dependent protein kinase (IC50 = 8 nM) and p60v-src (IC50 = 6 nM).
  • Purity
    > 98%
  • CAS Number
    62996-74-1
  • Chemical structure
    Chemical Structure

Properties

  • Chemical name
    [9S-(9α,10β,11β,13α)]-2,3,10,11,12,13-Hexahydro-10-methoxy-9-methyl-11-(methylamino)-9,13-epoxy-1H,9H-diindolo[1,2,3-gh:3,2,1-lm]pyrrolo[3,4-j][1,7]benzodiazonin-1-one
  • Molecular weight
    466.54
  • Molecular formula
    C28H26N4O3
  • PubChem identifier
    44259
  • Storage instructions
    Store at +4°C. Store under desiccating conditions. The product can be stored for up to 12 months.
  • Solubility overview
    Soluble in DMSO to 50 mM
  • Handling

    This product is supplied in one (or more) pack size which is freeze dried. Therefore the contents may not be readily visible, as they can coat the bottom or walls of the vial. Please see our FAQs and information page for more details on 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.

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

  • SMILES
    O=C6NCc5c2c4n(c1ccccc12)[C@@]3(C)O[C@H](C[C@@H](NC)[C@H]3OC)n7c4c(c56)c8ccccc78
  • Source

    Synthetic

  • Research areas

Images

  • HeLa and Jurkat cells were treated with 1 µM Staurosporine (STS) (ab120056) for 4 hours in complete cell culture media to induce apoptosis and cleaved PARP protein. Untreated and STS treated HeLa and Jurkat lysates were prepared in 1X Cell Extraction Buffer PTR and tested the Cleaved PARP SimpleStepTM ELISA. Raw OD 450 nm values are shown for 500 µg/mL lysate loads.
  • Lane 1: HeLa, vehicle (DMSO) treated for 4 hours Lane 2: HeLa 1 µM staurpsorine (ab120056), 4 hours Load 20 µg/lane 5% milk/PBST for block and antibody diluent Primary antibodies (2 hours, room temp) All lanes: ab136812 250X Primary Antibodies Cocktail, 1/250 dilution Secondary antibodies (1 hour, room temp) All lanes: ab136812 100X HRP-Conjugated Secondary Antibodies Cocktail, 1/100 dilution

  • Example of HeLa staurosporine (ab120056) treated cell lysate titration. Background-subtracted data values (mean +/- SD) are graphed.
  • Example of IC50 determination. HeLa cells were treated with a dose titration of Staurosporine for 4 hours in complete media. Cells were cultured and treated in a 96-well cell culture microtiter plate. Lysates were prepared by direct in-well lysis without media removal: 2X Cell Extraction Buffer PTR was added to an equal volume of media and then resulting lysate was used directly in the Cleaved PARP SimpleStepTM ELISA assay. Raw values for triplicate measurements are plotted. The calculated IC50 is 0.77 µM.

  • Demonstration of Cleaved PARP capture antibody specificity by western blot assay. 20 µg of HeLa extracts that were untreated or treated for 4 hours with 1 µM Staurosporine were analyzed by western blot. The GAPDH blot is included to show the relative loads of each lysate. In the HeLa cell line, Staurosporine treatment is required to detect cleaved PARP protein, as observed in the SimpleStep ELISA.

References

This product has been referenced in:
  • Gadbois DM  et al. Staurosporine is a potent inhibitor of p34cdc2 and p34cdc2-like kinases. Biochem Biophys Res Commun 184:80-5 (1992). Read more (PubMed: 1567459) »
  • Rüegg UT & Burgess GM Staurosporine, K-252 and UCN-01: potent but nonspecific inhibitors of protein kinases. Trends Pharmacol Sci 10:218-20 (1989). Read more (PubMed: 2672462) »
See all 5 Publications for this product

Customer reviews and Q&As

1-6 of 6 Q&A

Answer

Thank you for your response.

I would recommend preparing a 1 mM stock solution and using it at a 1 ul concentration in cell medium.



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.


This is to let you know that I have placed a new order for you - for one vial ofab120056 as a free of charge replacement and the new order number is 1075953.

I hope the second vial will work as it is expected and please do let me know how you are getting on with this product.

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Answer

Just to confirm that our Lab has finished a quality control (QC) analysis of the compound, which it has passed. We are therefore confident that the compound remains of high purity.

I can certainly offer you a replacement vial and would suggest that you run a positive control to confirm that the assay is running as it should be.

I hope this helps and if I can assist further, please do not hesitate to contact me.

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Answer

Thank you for getting back to me with some further details.

Our Lab is currently undertaking a quality control analysis on this batch of the compound to ensure that it remains of high purity. We expect a result back tomorrow although I expect that there won’t be anything wrong with the compound.

I can offer you a free of charge replacement vial but the vial will be from the same batch since we do not have new batch currently. I would advise you to ensure that a positive control is used to make sure that the assay is working correctly.

Let me know how you wish to proceed. I look forward to hearing from you soon.

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Answer

Thank you for the clarification and for your quick response.

It seems that the staurosporine is used at the right concentration for these cells so we are unsure why they are not detecting any apoptosis.

Would you be so kind to confirm how the cell viability has been checked and whether or not the cell look viable (i.e. example under a light microscope)?
Are you using a positive control for the assays?



I look forward to hearing from you soon.

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Answer

Thank you for getting back to me and for providing some further details.

I have discussed your enquiry with my colleagues in the Lab. Could you please send details of the calculations used to reconstitute the solution.

We are a bit confused regarding the concentrations stated. It may just be a typo but it is very unlikely to produce 200 uM from a 100 uM stock solution.

We are also wondering if this compound has been used at a concentration that may not be right. However, we have found an article that suggests that a lower concentration of Staurosporine is 10-7 M and a high concentration is 10-6 M. These concentrations fall within the range you have stated.

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

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Answer

Thank you for your enquiry and your interest in our products.

We have sold several vials of this product and not received any complaints. Therefore we are not aware of any-quality-related issue regarding ab120056.

It would be much appreciated if I could get some more information which would help me identify the source of the problem.

Q1: Have you carried out some time- and concentration course experiments with ab120056?

Q2: Are the cell still alive after treatment? Have you checked viability?

Q3: Could you please specify what sort of assay you are using to detect apoptotic cells i.e. Casapase (1, 2, 3, 8, 9), Annexin V, DNA ladder, TUNEL?

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

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

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