For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    products/sample-preparation-kits/hela-serthr-phosphatase-inhibited-whole-cell-lysate-set-calyculin-treated-and-vehicle-treated-ab135395.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Kits/ Lysates/ Other Lysates Membrane Lysates Human Muscle/Skin/Skeleton
Share by email

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

  • Datasheet
  • SDS
Submit a review Q&A (1)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Western Blot
  • SDS-PAGE

You may also be interested in

Lysate
Product image
HeLa whole cell lysate (ab150035)
Primary
Product image
Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)

View more associated products

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

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 2 units
    Calyculin-Treated HeLa Lysate 1 x 200µg
    Control for Calyculin-Treated HeLa Lysate 1 x 200µg
  • Research areas

    • Kits/ Lysates/ Other
    • Lysates
    • Membrane Lysates
    • Human
    • Muscle/Skin/Skeleton
    • Kits/ Lysates/ Other
    • Lysates
    • Cytoskeletal Lysates
    • Human
    • Muscle/Skin/Skeleton
    • Kits/ Lysates/ Other
    • Lysates
    • Nuclear Lysates
    • Human
    • Muscle/Skin/Skeleton
    • Kits/ Lysates/ Other
    • Lysates
    • Whole Cell Lysates
    • Human
    • Muscle/Skin/Skeleton

Associated products

  • Positive Controls

    • Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
    • Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Related Products

    • Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)

Applications

The Abpromise guarantee

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

Images

  • Western Blot
    Western Blot

    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.

  • SDS-PAGE
    SDS-PAGE
    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

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab135395? Please let us know so that we can cite the reference in this datasheet.

ab135395 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Question



We want to order this item (135395) as a control for some of our experiments. We are interested in the vehicle treated lysate only. However, I have a few technical questions before we place the order-

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)?

2. Are the cell debris (nuclei, mitochondria) removed from this? And are the lipids also removed?

3. Do you have any information about the cell-density of the HeLa cells used prior to the lysis?

Read More

Abcam community

Verified customer

Asked on May 26 2015

Answer



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. Since SDS generates a sticky lysate due to DNA content, the lysate is sonicated on ice to break the DNA. The formulation is 50mM Tris pH 6.8, 150mM NaCl, 8mM NaF, 1% SDS, 100mM DTT, plus protease inhibitor cocktail.



2. Are the cell debris (nuclei, mitochondria) removed from this? And are the lipids also removed?


- No lipids or cells debris is removed as this is a whole cell lysate.



3. Do you have any information about the cell-density of the HeLa cells used prior to the lysis?


- I can confirm that we generate the material from multiple plates that are about 95% confluent.



Read More

Anja Hoffmann

Abcam Scientific Support

Answered on May 26 2015

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Get resources and offers direct to your inbox Sign up
A-Z by research area
  • Cancer
  • Cardiovascular
  • Cell biology
  • Developmental biology
  • Epigenetics & Nuclear signaling
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal transduction
  • Stem cells
A-Z by product type
  • Primary antibodies
  • Secondary antibodies
  • Biochemicals
  • Isotype controls
  • Flow cytometry multi-color selector
  • Kits
  • Loading controls
  • Lysates
  • Peptides
  • Proteins
  • Slides
  • Tags and cell markers
  • Tools & Reagents
Help & support
  • Support
  • Make an Inquiry
  • Protocols & troubleshooting
  • Placing an order
  • RabMAb products
  • Biochemical product FAQs
  • Training
  • Browse by Target
Company
  • Corporate site
  • Investor relations
  • Company news
  • Careers
  • About us
  • Blog
Events
  • Tradeshows
  • Conferences
International websites
  • abcam.cn
  • abcam.co.jp

Join with us

  • LinkedIn
  • facebook
  • Twitter
  • YouTube
  • Terms of sale
  • Website terms of use
  • Cookie policy
  • Privacy policy
  • Legal
  • Modern slavery statement
© 1998-2023 Abcam plc. All rights reserved.