Overview

  • Product name
  • Description
    Rabbit polyclonal to LAMP2A
  • Tested applications
    Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human, Monkey, Chinese hamster
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide corresponding to Human LAMP2A aa 350 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). LAMP2 has 3 distinct isoforms, LAMP2A, 2B & 2C.
    Database link: P13473
    (Peptide available as ab23322)

  • Positive control
    • This antibody gave a positive signal in Western Blot in MEF1 whole cell lysate and the following tissue lysates: Mouse Lung, Human Liver, Human Liver (membrane fraction).

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
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab18528 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 Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 105 kDa (predicted molecular weight: 100 kDa).

Abcam recommends using 3% milk as the blocking agent.

ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval 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
    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
    • 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 images

  • ab18528 stained MEF1 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18528 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • Lanes 1 - 3 : Anti-LAMP2A antibody (ab18528) at 1 µg/ml
    Lanes 4 - 5 : non Abcam anti-LAMP2 antibody

    Lane 1 : MEF lysate extracted using 0.5 % NP40
    Lane 2 : HeLa lysate extracted using 1%Triton X-100
    Lane 3 : HeLa lysate extracted using 0.5 % NP40
    Lane 4 : MEF lysate extracted using 1% Triton X-100
    Lane 5 : MEF lysate extracted using 0.5 % NP40

    Secondary
    goat anti-rabbit at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size : 100 kDa

    This image and legend is taken from an Abreview by Eeva-Liisa Eskelinen submitted in December 2007

    Western blotting with the anti-LAMP2A (lanes 1-3) and an alternative antibody against the luminal domain of mouse LAMP2 (lanes 4 & 5) using MEF & HeLa cells extracted using either 1% Triton X-100 or 0.5 % NP40. Ab18528 recognized a 100 kDa band corresponding intact LAMP2 and also recognized mouse LAMP2 as expected from the sequence similarity. LAMP2 undergoes extensive glycosylation so ab18528 detects a band of ~ 100 kDa by Western blot.

  • IHC image of Lamp2A staining in human breast adenocarcinoma 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 ab18528, 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.

  • ab18528 staining Lamp2A in CaCO2 cells treated with SB 202190 (ab120638), by ICC/IF. Increase of Lamp2A expression correlates with increased concentration of SB 202190, as described in literature.
    The cells were incubated at 37°C for 3 hours in media containing different concentrations of ab120638 (SB 202190) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab18528 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • All lanes : Anti-LAMP2A antibody (ab18528) at 1 µg/ml (blocked with 3% Milk)

    Lane 1 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : Lung (Mouse) Tissue Lysate
    Lane 3 : Human liver tissue lysate - total protein (ab29889)
    Lane 4 : Human liver left lobe tissue lysate - membrane extract (ab29086)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 100 kDa
    Observed band size : 105 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 30 kDa,35 kDa,55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes

    Abcam recommends using 3% milk as the blocking agent.

  • ICC/IF image of ab18528 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18528, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • ICC/IF image of ab18528 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18528, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-LAMP2A antibody (ab18528)

This product has been referenced in:
  • Miles AL  et al. The vacuolar-ATPase complex and assembly factors, TMEM199 and CCDC115, control HIF1a prolyl hydroxylation by regulating cellular iron levels. Elife 6:N/A (2017). WB ; Human . Read more (PubMed: 28296633) »
  • Cao H  et al. The Endocytic Fate of the Transferrin Receptor Is Regulated by c-Abl Kinase. J Biol Chem 291:16424-37 (2016). Read more (PubMed: 27226592) »

See all 28 Publications 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 (U2OS cells)
Gel Running Conditions
Reduced Denaturing (3-8% Gradient Gel)
Loading amount
50 µg
Specification
U2OS cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 25 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Tissue lysate - whole (mouse heart)
Total protein in input
200 µg
Immuno-precipitation step
Other - Dynabeads IgG
Specification
mouse heart
Username

Abcam user community

Verified customer

Submitted Feb 08 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Chinese Hamster Cell (CHO cell)
Specification
CHO cell
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 18 2015

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing
Sample
Chinese Hamster Cell lysate - whole cell (CHO cell)
Specification
CHO cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Username

Abcam user community

Verified customer

Submitted Nov 03 2014

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (4-12% Nupage gel)
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Oct 03 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (HUVEC)
Specification
HUVEC
Permeabilization
Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
Fixative
Paraformaldehyde
Username

Dr. Dimitra Kalamida

Verified customer

Submitted Nov 21 2013

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 (Cortical tissue)
Specification
Cortical tissue
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

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (FRhK-4 cells, kidney)
Specification
FRhK-4 cells, kidney
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,2%Tx-100 in PBS
Username

Abcam user community

Verified customer

Submitted Apr 02 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Monkey Cell lysate - whole cell (FRhK-4 cells. kidney)
Loading amount
50 µg
Specification
FRhK-4 cells. kidney
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Abcam user community

Verified customer

Submitted Apr 02 2013

Thank you for contacting us.

I have checked our catalog for Lamp2 antibodies that have been tested in IP, and we have only the following 2 antibodies:

ab13524: Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (tested IP, human; rat mo...

Read More

1-10 of 16 Abreviews or Q&A

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

Sign up