Recombinant Anti-LAMP2 antibody [EPR19531] (ab199947)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19531] to LAMP2
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-LAMP2 antibody [EPR19531]
See all LAMP2 primary antibodies -
Description
Rabbit monoclonal [EPR19531] to LAMP2 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal kidney, fetal spleen and placenta lysates; HeLa, THP-1, HepG2, HEK-293, JAR and Jurkat whole cell lysates. IHC-P: Human liver and breast cancer tissues.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19531 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab199947 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 |
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Detects a band of approximately 110-130 kDa (predicted molecular weight: 45 kDa).
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Notes |
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IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 110-130 kDa (predicted molecular weight: 45 kDa). |
Target
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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
Defects in LAMP2 are the cause of Danon disease (DAND) [MIM:300257]; also known as glycogen storage disease type 2B (GSD2B). DAND is a lysosomal glycogen storage disease characterized by the clinical triad of cardiomyopathy, vacuolar myopathy and mental retardation. It is often associated with an accumulation of glycogen in muscle and lysosomes. -
Sequence similarities
Belongs to the LAMP family. -
Post-translational
modificationsO- 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
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Database links
- Entrez Gene: 3920 Human
- Omim: 309060 Human
- SwissProt: P13473 Human
- Unigene: 496684 Human
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Form
Alternative splicing produces 3 isoforms. -
Alternative names
- CD107 antigen like family member B antibody
- CD107 antigen-like family member B antibody
- CD107b antibody
see all
Images
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All lanes : Anti-LAMP2 antibody [EPR19531] (ab199947) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 cell lysate
Lane 2 : LAMP2 knockout HEK-293 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : LAMP2 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 45 kDaLanes 1 - 4: Merged signal (red and green). Green - ab199947 observed at 110 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab199947 was shown to react with LAMP2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255402 (knockout cell lysate ab263861) was used. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. ab199947 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-LAMP2 antibody [EPR19531] (ab199947) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : LAMP2 knockout HEK-293 whole cell lysate
Lane 3 : HepG2 whole cell lysate
Lane 4 : THP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 45 kDaLanes 1 - 4: Merged signal (red and green). Green - ab199947 observed at 45 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab199947 was shown to specifically react with LAMP2 in wild-type HEK-293 cells as signal was lost in LAMP2 knockout cells. Wild-type and LAMP2 knockout samples were subjected to SDS-PAGE. Ab199947 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-LAMP2 antibody [EPR19531] (ab199947) at 1/1000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal spleen lysate
Lane 3 : Human placenta lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution
Predicted band size: 45 kDa
Observed band size: 110-130 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 19828315) and lots of other products.
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All lanes : Anti-LAMP2 antibody [EPR19531] (ab199947) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : THP-1 (Human monocytic leukemia cell line) whole cell lysate
Lane 3 : JAR (Human placenta choriocarcinoma cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 45 kDa
Observed band size: 110-130 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 19828315) and lots of other products.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19531] (ab199947)
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling LAMP2 with ab199947 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on Human liver 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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [EPR19531] (ab199947)
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling LAMP2 with ab199947 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on Human breast cancer 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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab199947 has been referenced in 1 publication.
- Zhang A et al. Lactate-induced M2 polarization of tumor-associated macrophages promotes the invasion of pituitary adenoma by secreting CCL17. Theranostics 11:3839-3852 (2021). PubMed: 33664865