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    products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299.pdf

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Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)

  • Datasheet
  • SDS
  • COA
Reviews (1)Q&A (2)References (5)

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Chemical Structure - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Immunocytochemistry/ Immunofluorescence - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Immunocytochemistry - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)

Key features and details

  • Inhibitor of ADP-ribosylation factor
  • CAS Number: 20350-15-6
  • Soluble in DMSO to 50 mM
  • Form / State: Solid
  • Source: Eupenicillium brefeldianum

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Overview

  • Product name

    Brefeldin A, Inhibitor of ADP-ribosylation factor
  • Description

    Inhibitor of ADP-ribosylation factor
  • Alternative names

    • Ascotoxin
    • BFA
    • Cyanein
    • Decumbin
  • Biological description

    Reversible inhibitor of protein translocation from the endoplasmic reticulum to the Golgi apparatus. Inhibits binding of the cytosolic coat protein, β-COP and ADP-ribosylation factor (ARF) to Golgi membranes and inhibits GDP-GTP exchange.

  • CAS Number

    20350-15-6
  • Chemical structure

    Chemical Structure

Properties

  • Chemical name

    (1R,2E,6S,10E,11aS,13S,14aR)-1,13-Dihydroxy-6-methyl-1,6,7,8,9,11a,12,13,14,14a-decahydro-4H-cyclopenta[f]oxacyclotridecin-4-one
  • Molecular weight

    280.36
  • Molecular formula

    C16H24O4
  • PubChem identifier

    5287620
  • Storage instructions

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

    Soluble in DMSO to 50 mM
  • 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[C@@H]1C[C@H]2[C@H](O)C=CC(=O)O[C@@H](C)CCCC=C[C@@H]2C1
  • Source

    Eupenicillium brefeldianum

  • Research areas

    • Microbiology
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Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab120299 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
Functional Studies
Use at an assay dependent concentration.
Notes
Functional Studies
Use at an assay dependent concentration.

Images

  • Chemical Structure - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Chemical Structure - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    2D chemical structure image of ab120299, Brefeldin A, Inhibitor of ADP-ribosylation factor
  • Immunocytochemistry/ Immunofluorescence - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Immunocytochemistry/ Immunofluorescence - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    ab84340 staining golgin-97 in MCF7 cells treated with brefeldin A (ab120299), by ICC/IF. Increase in Golgin-97 expression correlates with increased concentration of brefeldin A, as described in literature.
    The cells were incubated at 37°C for 1.5 h in media containing different concentrations of ab120299 (brefeldin A) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab84340 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    All lanes : Anti-MCP3 antibody [EPR22649-155] (ab228979) at 1/1000 dilution

    Lane 1 : Wild-type RAW 264.7 untreated control cell lysate
    Lane 2 : Wild-type RAW 264.7 PMA treated (80 nM, 24 h) plus LPS treated (100 ng/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
    Lane 3 : CCL7/MCP3 knockout RAW 264.7 untreated cell lysate
    Lane 4 : CCL7/MCP3 knockout RAW 264.7 PMA treated (80 nM, 24 h) plus LPS treated (100 ng/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
    Lane 5 : J774A.1 Glucose treated (138.8 mMol/L, 8 h) plus Brefeldin A treated (5 µg/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate
    Lane 6 : J774A.1 Glucose treated (5.6 mMol/L, 8 h) plus Brefeldin A treated (5 µg/ml, 6 h) and Brefeldin A (ab120299) treated (5 µg/ml, 5 h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Observed band size: 14 kDa why is the actual band size different from the predicted?



    Lanes 1 - 6: Merged signal (red and green). Green - ab228979 observed at 14 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab228979 was shown to react with MCP3 in RAW 264.7 wild-type cells in Western blot with loss of signal observed in CCL7 knockout sample. Wild-type and CCL7 knockout RAW 264.7 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab228979 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    All lanes : Anti-TNF alpha antibody [EPR22598-212] (ab255275) at 1/1000 dilution

    Lane 1 : Wild-type THP-1 Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 2 : Wild-type THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 3 : TNF alpha knockout THP-1 Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 4 : TNF alpha knockout THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 5 : U937 PMA treated (10 mM, 2 days) plus 16 h no treatment and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 6 : U937 PMA treated (10 mM, 2 days) and LPS treated (1 µg/ml, 16 h) plus Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Observed band size: 26 kDa why is the actual band size different from the predicted?



    Lanes 1 - 6: Merged signal (red and green). Green - ab255275 observed at 26 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab255275 was shown to react with TNF alpha in THP-1 wild-type cells in Western blot with loss of signal observed in TNF knockout sample. Wild-type and TNF knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab255275 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    All lanes : Anti-TNF alpha antibody [EPR19147] (ab183218) at 1/1000 dilution

    Lane 1 : Wild-type THP-1 Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 2 : Wild-type THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 3 : TNF alpha knockout THP-1 Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 4 : TNF alpha knockout THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 5 : U937 PMA treated (10 mM, 2 days) plus 16 h no treatment and Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate
    Lane 6 : U937 PMA treated (10 mM, 2 days) and LPS treated (1 µg/ml, 16 h) plus Brefeldin A (ab120299) treated (5 µg/ml, 4 h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Observed band size: 26 kDa why is the actual band size different from the predicted?



    Lanes 1 - 6: Merged signal (red and green). Green - ab183218 observed at 26 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab183218 was shown to react with TNF alpha in THP-1 wild-type cells in Western blot with loss of signal observed in TNF knockout sample. Wild-type and TNF knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab183218 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Immunocytochemistry - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)

    ab66579 staining TNF alpha in Raw264.7 cells. The cells were treated with LPS for 7 hours and Brefeldin A (ab120299) for the final 3 hours. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab66579 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

     

    Also suitable in cells fixed with 4% paraformaldehyde (10 min).

     

    Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    All lanes : Anti-IP10 antibody [EPR20764] (ab214668) at 1/1000 dilution

    Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate
    Lane 2 : Wild-type A549 IFN-y (ab259377) (100 ng/ml, 32 h) and TNF-alpha (ab259410) (10 ng/ml, 32h), and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate
    Lane 3 : IP10 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate
    Lane 4 : IP10 knockout A549 IFN-y (ab259377) (100ng/ml, 32h) and TNF-alpha (ab259410) (10ng/ml, 32h), and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate
    Lane 5 : THP-1 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate
    Lane 6 : THP-1 IFN-y (ab259377) (200ng/ml, 24h) and LPS (50ng/ml, 24h)-treated for 24 hours, and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Observed band size: 11 kDa why is the actual band size different from the predicted?



    Lanes 1 - 6: Merged signal (red and green). Green - ab214668 observed at 11 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab214668 was shown to react with IP10 in wild-type A549 cells in western blot with loss of signal observed in IP10 knockout cell line ab266971 (knockout cell lysate ab256888). Wild-type and IP10 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab214668 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    All lanes : Anti-IP10 antibody [EPR7850] (ab137018) at 1/500 dilution

    Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate
    Lane 2 : Wild-type A549 IFN-y (ab259377) (100 ng/ml, 32 h) and TNF-alpha (ab259410) (10 ng/ml) for 32 hours, and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate
    Lane 3 : IP10 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate
    Lane 4 : IP10 knockout A549 IFN-y (ab259377) (100 ng/ml, 32 h) and TNF-alpha (ab259410) (10 ng/ml) for 32 hours, and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Observed band size: 11 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab137018 observed at 11 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab137018 was shown to react with IP10 in A549 wild-type cells in western blot with loss of signal observed in IP10 knockout cell line ab266969 (IP10 knockout cell lysate ab256886). A549 wild-type and IP10 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab137018 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    All lanes : Anti-IL-6 antibody [EPR21711] (ab233706) at 1/1000 dilution

    Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate
    Lane 2 : Wild-type A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate
    Lane 3 : IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate
    Lane 4 : IL-6 knockout A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Observed band size: 25 kDa why is the actual band size different from the predicted?
    Additional bands at: 40 kDa (possible non-specific binding)



    Lanes 1 - 4: Merged signal (red and green). Green - ab233706 observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

    ab233706 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type and IL-6 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab233706 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    Western blot - Brefeldin A, Inhibitor of ADP-ribosylation factor (ab120299)
    All lanes : Anti-IL-6 antibody [EPR20653] (ab214429) at 1/1000 dilution

    Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate
    Lane 2 : Wild-type A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate
    Lane 3 : IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate
    Lane 4 : IL-6 knockout A549 IL-1ß (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.


    Lanes 1 - 4: Merged signal (red and green). Green - ab214429 observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

    ab214429 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type A549 and IL-6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab214429 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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
  • COA

References (5)

Publishing research using ab120299? Please let us know so that we can cite the reference in this datasheet.

ab120299 has been referenced in 5 publications.

  • Chen X  et al. Oxidative stress-induced IL-15 trans-presentation in keratinocytes contributes to CD8+ T cells activation via JAK-STAT pathway in vitiligo. Free Radic Biol Med 139:80-91 (2019). PubMed: 31078730
  • Ioannou MS  et al. Neuron-Astrocyte Metabolic Coupling Protects against Activity-Induced Fatty Acid Toxicity. Cell 177:1522-1535.e14 (2019). PubMed: 31130380
  • Martínez-Muñoz L  et al. Separating Actin-Dependent Chemokine Receptor Nanoclustering from Dimerization Indicates a Role for Clustering in CXCR4 Signaling and Function. Mol Cell 70:106-119.e10 (2018). PubMed: 29625032
  • Saxena S  et al. Neuroprotection through excitability and mTOR required in ALS motoneurons to delay disease and extend survival. Neuron 80:80-96 (2013). PubMed: 24094105
  • Pryazhnikov E  et al. Unusually Strong Temperature Dependence of P2X3 Receptor Traffic to the Plasma Membrane. Front Cell Neurosci 5:27 (2011). PubMed: 22194716

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Efficient inhibition of ER to Golgi translocation

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Sub-cellular frationation followed by WB of the protein of interest confirmed reduction of ER to Golgi translocation of the protein of interest. Incubated with 10ug/ml of Brefeldin A for 4 hours.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 29 2017

Question

I've purchased your Brefeldin A (ab120299) and I' ve used the DMSO to dissolve the powder to prepare some aliquots.

These aliquots have been stored at-20°C;

My question is: where can I store the aliquote once thawed to preseve the power of the drug?

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Abcam community

Verified customer

Asked on Jun 09 2015

Answer

We would always recommend storing the reconstituted product at -20ºC until it is ready for use as other suppliers also suggest. Generally, these aliquots will be useable for up to one month. Some customers still use the product for up to 3 months but longer may increase the loss of potency.

Once thawed we would recommend using it immediately and have not tested any other storage conditions.

Having done a literature search, it seems like some researchers store it at 4ºC whilst they do their different experiments. Please note however that DMSO freezes at this temperature and so the solution needs to be completely thawed each time. This is why we would not recommend doing this often as every time you undergo a freeze thaw cycle you decrease the quality of the product so this should be kept to a minimum and experiments planned accordingly to try and decrease the number of freeze thaw cycles with one aliquot.

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Elisa Thomas

Abcam Scientific Support

Answered on Jun 09 2015

Question

DearAll,
could you please let me know if ab11458, ab95536, ab47228can be used for intracytoplasmic stainingin FACS analysis?

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Abcam community

Verified customer

Asked on Feb 28 2012

Answer

Thank you for contacting us.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase. For the mentioned TNF alpha antibodies I can confirm that they can be used for intracytoplasmic staining in flow cytometry.

Please note that ab11458 requires membrane permeabilisation, as mentioned on the datasheet. And in order to prevent secretion of the cytokine(s), an inhibitor such as Brefeldin A (ab120299) is recommended.

ab120299: Click here (or use the following: https://www.abcam.com/Brefeldin-A-Inhibitor-of-ADP-ribosylation-factor-ab120299.html).

We like to encourage you to submit an Abreview about one of the antibodies via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates. To find out more about our Abreview system, please see the following link: https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Abcam Scientific Support

Answered on Feb 28 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES, NOT FOR USE IN HUMANS"
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