Recombinant Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4207(2)] to LAMP2A - Lysosome Marker
- Suitable for: Flow Cyt, WB, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker
See all LAMP2A primary antibodies -
Description
Rabbit monoclonal [EPR4207(2)] to LAMP2A - Lysosome Marker -
Host species
Rabbit -
Specificity
ab125068 only recognizes LAMP2a. -
Tested applications
Suitable for: Flow Cyt, 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. Flow Cyt: HeLa cells.
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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.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4207(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
- HeLa whole cell lysate (ab150035)
- Mouse kidney normal tissue lysate - total protein (ab29305)
- HeLa whole cell lysate (ab29545)
- HeLa membrane extract lysate (ab29547)
- Jurkat whole cell lysate (ab30128)
- Jurkat membrane extract lysate (ab30130)
- NIH 3T3 whole cell lysate (ab7179)
- RAW 264.7 whole cell lysate (ab7187)
- Mouse kidney tissue lysate - total protein (0 days) (ab7261)
- Mouse kidney tissue lysate (14 days) (ab7262)
- Jurkat whole cell lysate (ab7899)
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Related Products
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 |
---|---|---|
Flow Cyt | 1/1000. | |
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. |
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
Danon disease -
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
- Entrez Gene: 16784 Mouse
- Omim: 309060 Human
- SwissProt: P13473 Human
- SwissProt: P17047 Mouse
- SwissProt: P17046 Rat
- Unigene: 496684 Human
- Unigene: 414016 Mouse
see all -
Form
Alternative splicing produces 3 isoforms. -
Alternative names
- CD107 antigen-like family member B antibody
- CD107b antibody
- LAMP 2 antibody
see all
Images
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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
All lanes : 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
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.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling LAMP2A (red) with ab125068 at a 1/1000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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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
All lanes : 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
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.
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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.
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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 minutesBlocking and Diluting buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
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.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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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.
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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
All lanes : 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 secondsBlocking and Diluting buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
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.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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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
All lanes : 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 minutesBlocking and Diluting buffer and concentration: 5% NFDM /TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2A antibody [EPR4207(2)] - Lysosome Marker (ab125068)
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.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
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
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 45 kDa
Exposure time: 3 minutesBlocking and Diluting buffer and concentration: 5% NFDM/TBST.
Protocols
Datasheets and documents
References (17)
ab125068 has been referenced in 17 publications.
- Liu X et al. PHLPP Sensitizes Multiple Myeloma Cells to Bortezomib Through Regulating LAMP2. Onco Targets Ther 13:401-411 (2020). PubMed: 32021285
- Francelle L et al. Inhibition of HDAC6 activity protects dopaminergic neurons from alpha-synuclein toxicity. Sci Rep 10:6064 (2020). PubMed: 32269243
- Bajbouj K et al. Estrogen Signaling Induces Mitochondrial Dysfunction-Associated Autophagy and Senescence in Breast Cancer Cells. Biology (Basel) 9:N/A (2020). PubMed: 32244623
- Liu DX et al. Exosomes derived from HBV-associated liver cancer promote chemoresistance by upregulating chaperone-mediated autophagy. Oncol Lett 17:323-331 (2019). PubMed: 30655770
- Wang R et al. Tumor cells induce LAMP2a expression in tumor-associated macrophage for cancer progression. EBioMedicine 40:118-134 (2019). PubMed: 30711520