Key features and details
- Rabbit polyclonal to LAMP2A - Lysosome Marker
- Suitable for: WB, ICC/IF, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-LAMP2A antibody - Lysosome Marker
See all LAMP2A primary antibodies
DescriptionRabbit polyclonal to LAMP2A - Lysosome Marker
Tested applicationsSuitable for: WB, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Dog, Monkey, Chinese hamster
- WB: Human liver tissue lysate. ICC: CaCo2 cells, HeLa and HepG2 cells. IHC-P: Human breast adenocarcinoma tissue.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
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.
|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.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
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.
modificationsO- 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
FormAlternative splicing produces 3 isoforms.
- CD107 antigen-like family member B antibody
- CD107b antibody
- LAMP 2 antibody
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.
IHC image of Lamp2A staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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.
All lanes : Anti-LAMP2A antibody - Lysosome Marker (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.
All lanes : 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.
Lanes 3 and 4 are human liver tissue lysates, total protein.
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.
ab18528 has been referenced in 98 publications.
- di Domenico A et al. Patient-Specific iPSC-Derived Astrocytes Contribute to Non-Cell-Autonomous Neurodegeneration in Parkinson's Disease. Stem Cell Reports N/A:N/A (2019). PubMed: 30639209
- Hoffmann AC et al. Extracellular aggregated alpha synuclein primarily triggers lysosomal dysfunction in neural cells prevented by trehalose. Sci Rep 9:544 (2019). PubMed: 30679445
- Wang R et al. Tumor cells induce LAMP2a expression in tumor-associated macrophage for cancer progression. EBioMedicine 40:118-134 (2019). PubMed: 30711520
- Zhang J et al. Chaperone-Mediated Autophagy Upregulation Rescues Megalin Expression and Localization in Cystinotic Proximal Tubule Cells. Front Endocrinol (Lausanne) 10:21 (2019). PubMed: 30774622
- Saito T et al. An alternative mitophagy pathway mediated by Rab9 protects the heart against ischemia. J Clin Invest 129:802-819 (2019). PubMed: 30511961