THP-1 nuclear extract lysate (PMA stimulated) (ab14901)

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Functional Studies - THP1 nuclear extract lysate (PMA stimulated) (ab14901)

    Overview

    • Product name

      THP-1 nuclear extract lysate (PMA stimulated)
      See all THP1 lysates

    • Description

      THP-1 nuclear extract lysate

    • General notes

      Cell line: THP1 (Human acute monocytic leukemia).
      The protein content has been determined by a Bradford-based assay.
      The THP-1 nuclear extracts have been collected in Lysis Buffer after a 24-hour incubation with or without PMA (10 ng/ml).

    Properties

    • Mycoplasma free

      Yes

    • Form

      Liquid

    • Storage instructions

      Shipped on Dry Ice. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.

    • Storage buffer

      pH: 7.50
      Constituents: 0.0002% Sodium molybdate, 0.216% Beta glycerophosphate, 0.018% Sodium orthovanadate, 0.042% Sodium fluoride, 0.015% DTT, 0.48% HEPES, 0.003% EDTA, 20% Glycerol, 2.34% Sodium chloride

      With Protease inhibitors.
    • Concentration information loading...

    • Research areas

    • Background

      Properties: differentiation; lysozyme synthesis; phagocytosis; Fc receptor; IL-1 prod.; C3b receptor. The cell line can differentiate into macrophage-like cells (phagocytic) and can be used for induction of differentiation studies. When stimulated with PMA cells have been reported to differentialte into a monocytic pathway and release arachidonic acid and prostanoids.

    Images

    • Functional Studies - THP1 nuclear extract lysate (PMA stimulated) (ab14901)
      Functional Studies - THP1 nuclear extract lysate (PMA stimulated) (ab14901)

      Each lot has been tested for PPAR-gamma activation by using TransAM™ PPAR-gamma Kits. The signal intensity for PPAR-gamma activation in each lot is compared to the signal intensity obtained with extracts from unstimulated THP-1 cells (see figure). After the signals are blanked, the ratio of the signals from stimulated cells over unstimulated cells must be above 4. This ratio may vary depending on the basal level of PPAR-gamma activation in a given cell type.

    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

    References (0)

    Publishing research using ab14901? Please let us know so that we can cite the reference in this datasheet.

    ab14901 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    Question

    J'ai oublié de mettre un anticorps anti PPAR gamma sur le bon de commande. jaurais besoin de l'avis de vos services tachniques : quel anticorps est il souhaitable de prendre pour du WB sur des lymphocytes B humains ? Pourraient ils m'envoyer leur protocole d'activation des THP1 avec du PMA ?

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    Answer

    Thank you for your enquiry. For the stimulation of THP1 cells with PMA please perform the following; Grow THP-1 cells at a density range of 5-8 x 10exp5 c/ml., add PMA to a final concentration of 10 ng/ml to stimulate the cells. Do not add anything to unstimulated cells. Incubate for 24 hrs. After 24 hrs, most of the stimulated cells should be attached to the flask. Then harvest the cells in ice-cold PBS buffer containing phosphatases inhibitors. Remove the PBS and lyse the cells by osmotic shock using a hypotonic buffer. Collect the nuclei and lyse them with lysis buffer (high salt buffer containing proteases inhibitors). I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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