Overview

  • Product name

    Anti-LAMP1 antibody [EPR21026]
    See all LAMP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR21026] to LAMP1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse LAMP1 aa 100-350. The exact sequence is proprietary.
    Database link: P11438

  • Positive control

    • WB: Mouse lung, colon, kidney and liver lysates; Neuro-2a, RAW 264.7 and NIH/3T3 whole cell lysates. IHC-P: Mouse spleen and lung tissues. IHC-Fr: Mouse kidney tissue. ICC/IF: NIH/3T3 and Neuro-2a cells. Flow Cyt: Neuro-2a cells. IP: NIH/3T3 whole cell lysate.
  • 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 ab208943 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 1/1000. Detects a band of approximately 90-120 kDa (predicted molecular weight: 43 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/100.

Perform heat mediated antigen retrieval using sodium citrate buffer (pH 6.0).

Flow Cyt 1/500.
IP 1/30.
ICC/IF 1/100.

Target

  • Function

    Presents carbohydrate ligands to selectins. Also implicated in tumor cell metastasis.
  • Sequence similarities

    Belongs to the LAMP family.
  • Post-translational
    modifications

    O- and N-glycosylated; some of the 18 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

  • Alternative names

    • CD107 antigen like family member A antibody
    • CD107 antigen-like family member A antibody
    • CD107a antibody
    • CD107a antigen antibody
    • LAMP 1 antibody
    • LAMP-1 antibody
    • LAMP1 antibody
    • LAMP1_HUMAN antibody
    • LAMPA antibody
    • LGP120 antibody
    • lgpA antibody
    • Lysosomal membrane glycoprotein 120KD antibody
    • Lysosomal Associated Membrane Protein 1 antibody
    • Lysosome associated membrane glycoprotein 1 antibody
    • Lysosome-associated membrane glycoprotein 1 antibody
    • Lysosome-associated membrane protein 1 antibody
    • OTTHUMP00000040663 antibody
    see all

Images

  • Immunofluorescent analysis of 100% methanol-fixed Neuro-2a (mouse neuroblastoma cell line) cells labeling LAMP1 with ab208943 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Neuro-2a cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889), at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methonol-permeabilized Neuro-2a (mouse neuroblastoma cell line) cell line labeling LAMP1 with ab208943 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

  • All lanes : Anti-LAMP1 antibody [EPR21026] (ab208943) at 1/2000 dilution

    Lane 1 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate at 20 µg
    Lane 2 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg
    Lane 3 : Mouse kidney lysate at 20 µg
    Lane 4 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg

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

    Developed using the ECL technique.

    Predicted band size: 43 kDa
    Observed band size: 90-120 kDa
    why is the actual band size different from the predicted?



    Exposure time : Lanes 1-2: 3 minutes; Lane 3: 5 seconds; Lane 4: 10 seconds.

    Blocking and dilution buffer: 5% NFDM/TBST.

     

    The varying band sizes are due to different levels of glycosylation (PMID: 10212251, PMID: 26246576).

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling LAMP1 with ab208943 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular cytoplasmic staining on mouse spleen was observed, performed on a Leica Biosystems BOND® RX instrument (PMID: 22008915). Counter stained with hematoxylin.
     
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
     
    Note: BOND® is a registered trademark of Leica Biosystems Melbourne Pty Ltd.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunofluorescent analysis of 100% methanol-fixed NIH/3T3 (mouse embryo fibroblast cell line) cells labeling LAMP1 with ab208943 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line.
     
    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889), at 1/200 dilution (red).
     
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling LAMP1 with ab208943 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular cytoplasmic staining on mouse spleen was observed, performed on a Leica Biosystems BOND® RX instrument (PMID: 22008915). Counter stained with hematoxylin.
     
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
     
    Note: BOND® is a registered trademark of Leica Biosystems Melbourne Pty Ltd.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse kidney tissue labeling LAMP1 with ab208943 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Cytoplasmic staining in the endothelial cells of glomeruli and epithelial cells of renal tubules (PMID: 23229015; PMID:23635510). The nuclear counter stain is DAPI (blue).
     
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
  • All lanes : Anti-LAMP1 antibody [EPR21026] (ab208943) at 1/1000 dilution

    Lane 1 : Mouse lung lysate
    Lane 2 : Mouse colon lysate
    Lane 3 : Mouse liver lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    Predicted band size: 43 kDa
    Observed band size: 90-120 kDa why is the actual band size different from the predicted?




    Exposure time : Lanes 1-2: 15 seconds; Lane 3: 5 seconds.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • LAMP1 was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab208943 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab208943 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10 μg (Input).

    Lane 2: ab208943 IP in NIH/3T3 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab208943 in NIH/3T3 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

References

ab208943 has not yet been referenced specifically in any publications.

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