Autophagy Marker (APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, LC3B) Antibody Sampler Panel (ab228525)

Overview

  • Product name
    Autophagy Marker (APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, LC3B) Antibody Sampler Panel
  • Product overview

    Autophagy Antibody Panel Sampler Panel (ab228525) contains multiple trial-sized versions of clones against APG5L/ATG5, ATG16L1, ATG4B, ATG9A, Beclin 1, and LC3B specifically selected for high performance in various applications. This panel contains 6 recombinant rabbit monoclonal antibodies, 5 of which are knock-out validated. They are provided as a sampler panel to allow you to easily evaluate each in your required applications. 


    For guidelines on how to use each antibody within the panel, please consult the individual datasheet for each antibody.


    Panel contains:


    - Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5 (20 µL) ab108327


    - Rabbit monoclonal [EPR15638] to ATG16L1 - N-terminal (20 µL) ab187671


    - Rabbit monoclonal [EPR6436(2)] to ATG4B (20 µL) ab154843


    - Rabbit monoclonal [EPR2450(2)] to ATG9A (20 µL) ab108338


    - Rabbit monoclonal [EPR19662] to Beclin 1 (20 µL) ab207612


    - Rabbit monoclonal [EPR18709] to LC3B (20 µL) ab192890

  • Notes

    Explore our range of antibody sample panels designed to provide you with a variety of trial-size antibodies in a convenient and cost-effective format.

    Directly conjugated versions of our antibodies are available and ready to use for multicolor flow cytometry or immunocytochemistry analysis. Carrier-free formulations are also available for easy conjugation to labels of your choice. Please refer to the ‘Associated products’ section below.

Properties

Images

  • ab192890 staining LC3B in HeLa cells +/- Chloroquine (50μM, 24 hours). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 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 ab192890 at 1μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ab108338, at 1/100, staining ATG9A in paraffin-embedded Human colon tissue by Immunohistochemistry.

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Beclin 1 with ab207612 at 1/200 dilution. Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton-X100 in PBS. Samples were incubated with primary antibody (1/200 in PBS) for 16 hours at 22°C. ab150081, a goat anti-rabbit IgG H + L  (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution.

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ATG4B knockout HAP1 cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab154843 observed at 47 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab154843 was shown to specifically react with ATG4B when ATG4B knockout samples were used. Wild-type and ATG4B knockout samples were subjected to SDS-PAGE. ab154843 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling ATG16L1 with ab187671 at 1/100 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: APG5L/ATG5 knockout HAP1 cell lysate (20 µg) 
    Lane 3: Raji cell lysate (20 µg)
    Lane 4: Jeg-3 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab108327 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab108327 was shown to specifically react with APG5L/ATG5 when APG5L/ATG5 knockout samples were used. Wild-type and APG5L/ATG5 knockout samples were subjected to SDS-PAGE. ab108327 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

References

ab228525 has not yet been referenced specifically in any publications.

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