Validated using a knockout cell line
Recombinant
RabMAb

Anti-AGL antibody [EPR8880] (ab133720)

Overview

  • Product name
    Anti-AGL antibody [EPR8880]
    See all AGL primary antibodies
  • Description
    Rabbit monoclonal [EPR8880] to AGL
  • Host species
    Rabbit
  • Specificity
    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human AGL aa 50-150.

  • Positive control
    • Human fetal muscle, mouse muscle, Human fetal heart, K562 and 293T cell lysates, Human liver tissue and Human muscle tissue, HeLa cells.
  • General notes

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133720 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
ICC/IF 1/100 - 1/250.
WB 1/1000 - 1/10000. Predicted molecular weight: 175 kDa.
IHC-P 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

For unpurified use at 1/100 - 1/250. 

Flow Cyt 1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/100-1/500.

 

Target

  • Function
    Multifunctional enzyme acting as 1,4-alpha-D-glucan:1,4-alpha-D-glucan 4-alpha-D-glycosyltransferase and amylo-1,6-glucosidase in glycogen degradation.
  • Tissue specificity
    Liver, kidney and lymphoblastoid cells express predominantly isoform 1; whereas muscle and heart express not only isoform 1, but also muscle-specific isoform mRNAs (isoforms 2, 3 and 4). Isoforms 5 and 6 are present in both liver and muscle.
  • Involvement in disease
    Glycogen storage disease 3
  • Sequence similarities
    Belongs to the glycogen debranching enzyme family.
  • Post-translational
    modifications
    The N-terminus is blocked.
    Ubiquitinated.
  • Cellular localization
    Cytoplasm. Under glycogenolytic conditions localizes to the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 1110061O17Rik antibody
    • 4-1 antibody
    • 4-glucantransferase antibody
    • 6-glucosidase antibody
    • 9430004C13Rik antibody
    • 9630046L06Rik antibody
    • AGL antibody
    • AI850929 antibody
    • Amylo 1 6 glucosidase 4 alpha glucanotransferase antibody
    • Amylo-1 antibody
    • Amylo-alpha-1 antibody
    • C77197 antibody
    • Dextrin 6-alpha-D-glucosidase antibody
    • GDE antibody
    • GDE_HUMAN antibody
    • Glycogen debrancher antibody
    • Glycogen debranching enzyme antibody
    • Glycogen storage disease type III antibody
    • Oligo-1 antibody
    see all

Images

  • Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling AGL with purified ab133720 at 1:80 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling AGL with Purified ab133720 at 1:100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor� 594) 1:200. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human skeletal muscle tissue sections labeling AGL with Purified ab133720 at 1:2000 dilution (0.4 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP)
    secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.

  • All lanes : Anti-AGL antibody [EPR8880] (ab133720) at 1/1000 dilution (purified)

    Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
    Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 175 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • All lanes : Anti-AGL antibody [EPR8880] (ab133720) at 1/5000 dilution (purified)

    Lane 1 : Human fetal muscle lysates
    Lane 2 : Human fetal heart lysates
    Lane 3 : Mouse muscle lysates
    Lane 4 : Rat muscle lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 175 kDa
    Observed band size: 175 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: AGL knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HepG2 whole cell lysate (20 µg)
    Lane 4: Hek293 whole cell lysate (20 µg
    Lanes 1 - 4: Merged signal (red and green). Green - ab133720 observed at 170 kDa. Red - loading control, ab18058, observed at 130 kDa.

    Unpurified ab133720 was shown to specifically react with AGL when AGL knockout samples were used. Wild-type and AGL knockout samples were subjected to SDS-PAGE.
    Unpurified Ab133720 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed 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 1/10000 dilution for 1 hour at room temperature before imaging.


  • All lanes : Anti-AGL antibody [EPR8880] (ab133720) at 1/1000 dilution (unpurified)

    Lane 1 : Human fetal muscle lysate
    Lane 2 : Mouse muscle lysate
    Lane 3 : Human fetal heart lysate
    Lane 4 : K562 cell lysate
    Lane 5 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 175 kDa

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling AGL with purified ab133720 at 1/250 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor®488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Immunohistochemical analysis of paraffin-embedded Human liver tissue labelled with unpurified ab133720 at 1/100 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human muscle tissue labelled with unpurified ab133720 at 1/100 dilution.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

ab133720 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab133720.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up