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    products/primary-antibodies/lamp2-antibody-epr19512-lysosome-marker-ab199946.pdf

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Tags & Cell Markers Subcellular Markers Organelles Lysosome
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
  • Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
  • Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
  • Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
  • Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19512] to LAMP2 - Lysosome Marker
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-LAMP2 antibody [EPR19512] - Lysosome Marker
    See all LAMP2 primary antibodies
  • Description

    Rabbit monoclonal [EPR19512] to LAMP2 - Lysosome Marker
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

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

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19512
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Lysosome
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Immunology
    • Secreted Molecules
    • Other secreted molecules
    • Cardiovascular
    • Heart
    • Autophagy
    • Autophagosome
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Myeloid Progenitor
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Monocytic Lineage
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • Autophagosome
    • Cancer
    • Cell Death
    • Autophagy
    • Autophagosome
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-LAMP2 antibody [EPR19512] - BSA and Azide free (ab240351)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human LAMP2 knockout HeLa cell line (ab255402)
  • KO cell lysates

    • Human LAMP2 knockout HeLa cell lysate (ab263861)
  • Recombinant Protein

    • Recombinant Human LAMP2 protein (ab114167)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab199946 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
WB
1/2000. Detects a band of approximately 110-130 kDa (predicted molecular weight: 45 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
1/2000. Detects a band of approximately 110-130 kDa (predicted molecular weight: 45 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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.
  • Target information above from: UniProt accession P13473 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3920 Human
    • Omim: 309060 Human
    • SwissProt: P13473 Human
    • Unigene: 496684 Human
    • 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 [EPR19512] - Lysosome Marker (ab199946)
      Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
      All lanes : Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946) at 1 µg/ml

      Lane 1 : Wild-type HEK-293 cell lysate
      Lane 2 : LAMP2 knockout HEK-293 cell lysate
      Lane 3 : Wild-type HeLa cell lysate
      Lane 4 : LAMP2 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 45 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab199946 observed at 110 kDa. Red - loading control, ab8245 observed at 37 kDa.

       ab199946 was shown to react with LAMP2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255402 (knockout cell lysate ab263861) was used. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. ab199946 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 µg/ml 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 [EPR19512] - Lysosome Marker (ab199946)
      Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
      All lanes : Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946) at 1/2000 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 - ab199946 observed at 45 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab199946 was shown to recognize LAMP2 in wild-type HEK-293 cells as signal was lost at the expected MW in LAMP2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab199946 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/2000 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)

      Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling LAMP2 with ab199946 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human kidney is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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

    • Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
      Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
      All lanes : Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946) at 1/2000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : Human fetal heart lysate
      Lane 3 : Human fetal kidney lysate
      Lane 4 : Human fetal spleen lysate
      Lane 5 : 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?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1 and 2: 15 seconds; Lane 3,4 and 5: 5 seconds.

      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 [EPR19512] - Lysosome Marker (ab199946)
      Western blot - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
      All lanes : Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946) at 1/2000 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?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1,2 and 3: 1 second; Lane 4: 5 seconds.

      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 [EPR19512] - Lysosome Marker (ab199946)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)

      Immunohistochemical analysis of paraffin-embedded Human thyroid cancer tissue labeling LAMP2 with ab199946 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human thyroid cancer is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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

    • Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)
      Anti-LAMP2 antibody [EPR19512] - Lysosome Marker (ab199946)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (3)

    Publishing research using ab199946? Please let us know so that we can cite the reference in this datasheet.

    ab199946 has been referenced in 3 publications.

    • Yang J  et al. Therapeutic Effects of Simultaneous Delivery of Nerve Growth Factor mRNA and Protein via Exosomes on Cerebral Ischemia. Mol Ther Nucleic Acids 21:512-522 (2020). PubMed: 32682291
    • Zhang L  et al. TSTA3 facilitates esophageal squamous cell carcinoma progression through regulating fucosylation of LAMP2 and ERBB2. Theranostics 10:11339-11358 (2020). PubMed: 33042286
    • Xiong YJ  et al. Enhancement of epithelial cell autophagy induced by sinensetin alleviates epithelial barrier dysfunction in colitis. Pharmacol Res 148:104461 (2019). PubMed: 31542404

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