Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

Overview

  • Product nameAnti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker
    See all LAMP2A primary antibodies
  • Description
    Rabbit monoclonal [EPR4207(2)] to LAMP2A - Lysosome Marker
  • Specificityab125068 only recognizes LAMP2a.
  • Tested applicationsSuitable for: WB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human LAMP2A aa 400 to the C-terminus.
    Database link: P13473-1

  • Positive control
    • WB: HeLa, Jurkat, ECV-304, JAR, NIH/3T3, RAW 264.7, PC-12 and C6 whole cell lysates. Mouse kidney lysates and Rat liver and kidney lysates. IHC-P: Human placenta and liver tissue. Mouse and rat liver tissue. IP: HeLa and RAW 264.7 whole cell lysates.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions available:

    Anti-LAMP2a antibody (Alexa Fluor® 488) [EPR4207(2)] - Lysosome Marker (ab184808)

Properties

Applications

Our Abpromise guarantee covers the use of ab125068 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 120 kDa (predicted molecular weight: 45 kDa).
IP 1/60 - 1/100.
IHC-P 1/200 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Target

  • FunctionImplicated 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 specificityIsoform 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 diseaseDanon disease
  • Sequence similaritiesBelongs to the LAMP family.
  • Post-translational
    modifications
    O- and N-glycosylated; some of the 16 N-linked glycans are polylactosaminoglycans.
  • Cellular localizationCell membrane. Endosome membrane. Lysosome membrane. This protein shuttles between lysosomes, endosomes, and the plasma membrane.
  • Information by UniProt
  • Database links
  • FormAlternative splicing produces 3 isoforms.
  • Alternative names
    • CD107 antigen-like family member B antibody
    • CD107b antibody
    • LAMP 2 antibody
    • Lamp 2a antibody
    • LAMP-2 antibody
    • LAMP2 antibody
    • LAMP2_HUMAN antibody
    • Lysosome-associated membrane glycoprotein 2 antibody
    • Lysosome-associated membrane protein 2 antibody
    see all

Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker images

  • All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/2000 dilution (purified)

    Lane 1 : Mouse kidney tissue lysate
    Lane 2 : Rat kidney tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 45 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

    Difference in MW may be caused by different degree of glycosylation.

  • All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/10000 dilution (purified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : ECV-304 cell lysate
    Lane 3 : JAR cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 45 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

    May be seen at ~50 kDa representing the unglycosylated isoforms of LAMP2 and ~120 kDa representing the glycosylated form.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling LAMP2A with purified ab125068 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • ab125068 (purified) at 1/60 immunoprecipitating LAMP2A in HeLa whole cell lysate.

    Lane 1 (input): HeLa whole cell lysate (10µg)

    Lane 2 (+): ab125068 + HeLa whole cell lysate (10µg).

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab125068 in HeLa whole cell lysate.

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/5000 dilution (unpurified) + HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 45 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

  • All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/5000 dilution (unpurified)

    Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 2 : ECV-304 (Human urinary bladder cancer cell line) whole cell lysate
    Lane 3 : JAR (Human placenta choriocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 45 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

  • All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/2000 dilution (unpurified)

    Lane 1 : Mouse kidney
    Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 45 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    Blocking and Diluting buffer and concentration: 5% NFDM /TBST

  • All lanes : Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068) at 1/2000 dilution (unpurified)

    Lane 1 : Rat liver
    Lane 2 : Rat kidney
    Lane 3 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 45 kDa


    Exposure time : 3 minutes

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

  • Immunohistochemical analysis of paraffin-embedded Human placenta labeling LAMP2A with unpurified ab125068 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on human placenta tissue 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.

  • Immunohistochemical analysis of paraffin-embedded Human liver labeling LAMP2A with unpurified ab125068 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on human liver tissue 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.

  • Immunohistochemical analysis of paraffin-embedded Mouse liver labeling LAMP2A with unpurified ab125068 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on mouse liver tissue 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.

  • Immunohistochemical analysis of paraffin-embedded Rat liver labeling LAMP2A with unpurified ab125068 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on rat liver tissue 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.

  • LAMP2A was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with unpurified ab125068 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab125068 at 1/2000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.

    Lane 1: HeLa whole cell lysate 10ug (Input).

    Lane 2: ab125068 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab125068 in HeLa whole cell lysate.

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

     

  • LAMP2A was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate with ab125068 at 1/100 dilution. Western blot was performed from the immunoprecipitate using unpurified ab125068 at 1/2000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.

    Lane 1: RAW 264.7 whole cell lysate 10ug (Input).

    Lane 2: ab125068 IP in RAW 264.7 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab125068 in RAW 264.7 whole cell lysate.

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

References for Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)

This product has been referenced in:
  • Wang B  et al. Essential control of mitochondrial morphology and function by chaperone-mediated autophagy through degradation of PARK7. Autophagy 12:1215-28 (2016). Read more (PubMed: 27171370) »

See 1 Publication for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions Reduced Denaturing
Loading amount 5 µg
Specification HEK293T
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Nov 28 2016

Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Sample Rat Tissue sections (brain)
Specification brain
Permeabilization Yes - triton-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 25 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Human Cell lysate - whole cell (BA9 - Cortical area)
Specification BA9 - Cortical area
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Dr. Martin Broadstock

Verified customer

Submitted Aug 19 2013

The IgG concentrations of the current lots are:

3768-1 (ab109534, lot GR68319) - 0.032 mg/ml

3660-1 (ab125068, lot GR75050) - 1.554 mg/ml

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