Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-LAMP2 antibody [EPR19531] (ab199947)

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Overview

  • Product name

    Anti-LAMP2 antibody [EPR19531]
    See all LAMP2 primary antibodies

  • Description

    Rabbit monoclonal [EPR19531] to LAMP2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human LAMP2 aa 50-150. The exact sequence is proprietary.
    Database link: P13473

  • Positive control

    • WB: Human fetal kidney, fetal spleen and placenta lysates; HeLa, THP-1, , HepG2, HEK-293, JAR and Jurkat whole cell lysates. IHC-P: Human liver and breast cancer tissues.

  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab199947 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
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 110-130 kDa (predicted molecular weight: 45 kDa).

Target

  • Function

    Implicated in tumor cell metastasis. May function in protection of the lysosomal membrane from autodigestion, maintenance of the acidic environment of the lysosome, adhesion when expressed on the cell surface (plasma membrane), and inter-and intracellular signal transduction. Protects cells from the toxic effects of methylating mutagens.

  • Tissue specificity

    Isoform LAMP-2A is highly expressed in placenta, lung and liver, less in kidney and pancreas, low in brain and skeletal muscle. Isoform LAMP-2B is highly expressed in skeletal muscle, less in brain, placenta, lung, kidney and pancreas, very low in liver.

  • Involvement in disease

    Defects in LAMP2 are the cause of Danon disease (DAND) [MIM:300257]; also known as glycogen storage disease type 2B (GSD2B). DAND is a lysosomal glycogen storage disease characterized by the clinical triad of cardiomyopathy, vacuolar myopathy and mental retardation. It is often associated with an accumulation of glycogen in muscle and lysosomes.

  • Sequence similarities

    Belongs to the LAMP family.

  • Post-translational
    modifications

    O- and N-glycosylated; some of the 16 N-linked glycans are polylactosaminoglycans.

  • Cellular localization

    Cell membrane. Endosome membrane. Lysosome membrane. This protein shuttles between lysosomes, endosomes, and the plasma membrane.
  • Information by UniProt

  • Database links

    • Form

      Alternative splicing produces 3 isoforms.

    • Alternative names

      • CD107 antigen like family member B antibody
      • CD107 antigen-like family member B antibody
      • CD107b antibody
      • LAMP 2 antibody
      • LAMP 2C antibody
      • LAMP-2 antibody
      • LAMP2 antibody
      • LAMP2_HUMAN antibody
      • LAMPB antibody
      • LGP 96 antibody
      • LGP110 antibody
      • LGP96 antibody
      • Lysosomal associated membrane protein 2 antibody
      • Lysosome associated membrane glycoprotein 2 antibody
      • Lysosome associated membrane protein 2 antibody
      • Lysosome-associated membrane glycoprotein 2 antibody
      • Lysosome-associated membrane protein 2 antibody
      • MAC3 antibody
      see all

    Images

    • Western blot - Anti-LAMP2 antibody [EPR19531] (ab199947)
      Western blot - Anti-LAMP2 antibody [EPR19531] (ab199947)
      Lane 1: HEK-293 wildtype cell lysate (20 µg)

      Lane 2: LAMP2 HEK-293 knockout cell lysate (20 µg)

      Lane 3: HeLa wildtype cell lysate (20 µg)

      Lane 4: LAMP2 HeLa knockout cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab199947 observed at 110 kDa. Red - loading control, ab8245 observed at 37 kDa.

      ab199947 was shown to react with LAMP2 in HeLa wildtype. Loss of signal was observed when knockout sample ab263861 was used. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. ab199947 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-LAMP2 antibody [EPR19531] (ab199947)
      Western blot - Anti-LAMP2 antibody [EPR19531] (ab199947)
      All lanes : Anti-LAMP2 antibody [EPR19531] (ab199947) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293 whole cell lysate
      Lane 2 : LAMP2 knockout HEK-293 whole cell lysate
      Lane 3 : HepG2 whole cell lysate
      Lane 4 : THP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 45 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab199947 observed at 45 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab199947 was shown to specifically react with LAMP2 in wild-type HEK-293 cells as signal was lost in LAMP2 knockout cells. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. Ab199947 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-LAMP2 antibody [EPR19531] (ab199947)
      Western blot - Anti-LAMP2 antibody [EPR19531] (ab199947)
      All lanes : Anti-LAMP2 antibody [EPR19531] (ab199947) at 1/1000 dilution

      Lane 1 : Human fetal kidney lysate
      Lane 2 : Human fetal spleen lysate
      Lane 3 : Human placenta lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

      Predicted band size: 45 kDa
      Observed band size: 110-130 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight  observed is consistent with what has been described in the literature (PMID: 19828315) and lots of other products.

    • Western blot - Anti-LAMP2 antibody [EPR19531] (ab199947)
      Western blot - Anti-LAMP2 antibody [EPR19531] (ab199947)
      All lanes : Anti-LAMP2 antibody [EPR19531] (ab199947) at 1/1000 dilution

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : THP-1 (Human monocytic leukemia cell line) whole cell lysate
      Lane 3 : JAR (Human placenta choriocarcinoma cell line) whole cell lysate
      Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 45 kDa
      Observed band size: 110-130 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight  observed is consistent with what has been described in the literature (PMID: 19828315) and lots of other products.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19531] (ab199947)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19531] (ab199947)

      Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling LAMP2 with ab199947 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Cytoplasm staining on Human liver is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19531] (ab199947)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19531] (ab199947)

      Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling LAMP2 with ab199947 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Cytoplasm staining on Human breast cancer is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    References

    ab199947 has not yet been referenced specifically in any publications.

    Publishing research using ab199947? Please let us know so that we can cite the reference in this datasheet.

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