Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15638] to ATG16L1 - BSA and Azide free
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free
    See all ATG16L1 primary antibodies

  • Description

    Rabbit monoclonal [EPR15638] to ATG16L1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, Jurkat, Daudi and HAP1 cell lysates. IHC-P: Human colon tissue.

  • General notes

    Ab232636 is the carrier-free version of ab187671. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab232636 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Images

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    All lanes : Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG16L1 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 68 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab187671).

      Lanes 1- 4: Merged signal (red and green). Green - ab187671 observed at 68 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab187671 was shown to react with ATG16L1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265263 (knockout cell lysate ab256842) was used. Wild-type HeLa and ATG16L1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)

    Immunohistochemical analysis of paraffin-embedded Human colon 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187671).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    All lanes : Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG16L1 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 68 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab187671).

    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab187671 Anti-ATG16L1 antibody [EPR15638] was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265263 (knockout cell lysate ab256842) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    All lanes : Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG16L1 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 68 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab187671).

    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 68 and 72 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab187671 Anti-ATG16L1 antibody [EPR15638] - N-terminal was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261773 (knockout cell lysate ab256844) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    All lanes : Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG16L1 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 68 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab187671).

    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 68 and 72 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab187671 Anti-ATG16L1 antibody [EPR15638] - N-terminal was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261772 (knockout cell lysate ab256843) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    Western blot - Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    All lanes : Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/2000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : ATG16L1 knockout HAP1 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 68 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab187671 observed at 68 and 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab187671 was shown to recognize ATG16L1 when ATG16L1 knockout samples were used, along with additional cross-reactive bands. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and ab8245 (loading control to GAPDH) were diluted 1/2000 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 h at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187671).

  • Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)
    Anti-ATG16L1 antibody [EPR15638] - BSA and Azide free (ab232636)

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