Anti-LAMP2B antibody - Lysosome Marker (ab18529)

Overview

  • Product name
    Anti-LAMP2B antibody - Lysosome Marker
    See all LAMP2B primary antibodies
  • Description
    Rabbit polyclonal to LAMP2B - Lysosome Marker
  • Specificity
    This product has low sequence homology with Mouse LAMP2B (67%), and high sequence homology with Mouse LAMP2C (91.7%).
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Cow, Human
  • Immunogen

    Synthetic peptide corresponding to Human LAMP2B aa 350 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). LAMP2 has 3 distinct isoforms, LAMP2A, 2B & 2C which are expressed in different tissues and differ in sequence at the extreme C-terminus.
    Database link: P13473
    (Peptide available as ab23321)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa, HEK293. This antibody gave a positive signal in the following Methanol fixed cell lines: HeLa. It also gave a positive signal in Human skin tissue sections.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab18529 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 Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 110 kDa).

Abcam recommends using milk as the blocking agent.

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
    Danon disease
  • 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
    • CD_antigen=CD107b antibody
    • CD107 antigen like family member B antibody
    • CD107 antigen-like family member B antibody
    • CD107b antibody
    • LAMP 2 antibody
    • Lamp 2b antibody
    • LAMP-2 antibody
    • LAMP2 antibody
    • LAMP2_HUMAN antibody
    • LAMPB antibody
    • LGP110 antibody
    • Lysosomal associated membrane protein 2 antibody
    • Lysosome associated membrane glycoprotein 2 antibody
    • Lysosome-associated membrane glycoprotein 2 antibody
    • Lysosome-associated membrane protein 2 antibody
    • MAC3 antibody
    see all

Anti-LAMP2B antibody - Lysosome Marker images

  • All lanes : Anti-LAMP2B antibody - Lysosome Marker (ab18529) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 110 kDa
    Observed band size : 110 kDa
    Additional bands at : 42 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab18529 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    LAMP2B undergoes extensive glycosylation so ab18529 detects a band of 110 kDa by Western blot.

  • ICC/IF image of ab18529 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18529, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • IHC image of ab18529 staining in human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18529, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-LAMP2B antibody - Lysosome Marker (ab18529)

This product has been referenced in:
  • Kibria G  et al. A rapid, automated surface protein profiling of single circulating exosomes in human blood. Sci Rep 6:36502 (2016). WB ; Human . Read more (PubMed: 27819324) »
  • Fujiwara N  et al. miR-634 Activates the Mitochondrial Apoptosis Pathway and Enhances Chemotherapy-Induced Cytotoxicity. Cancer Res 75:3890-901 (2015). Human . Read more (PubMed: 26216549) »

See all 10 Publications for this product

Product Wall

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 16 2013

Thank you very much for your interest in our products.

We do not offer free or trial sizes of our products simply because we carry over 100,000 items and having multiple sizes would be logistically impossible with our automated freezers. Read More

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Cow Cell (endothelial cells)
Specification
endothelial cells
Fixative
acetone/methanol
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Sep 17 2008

Application
Western blot
Sample
Mouse Cell lysate - whole cell (embryonic fibriblast)
Loading amount
50 µg
Specification
embryonic fibriblast
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Username

Dr. Eeva-Liisa Eskelinen

Verified customer

Submitted Dec 11 2007

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (cultured hippocampal astrocytes)
Specification
cultured hippocampal astrocytes
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.25% Triton X-100 for 5 min
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Aug 16 2007

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

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