Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)
Key features and details
- Mouse monoclonal [H4B4] to LAMP2 - Lysosome Marker
- Suitable for: ICC, ICC/IF, IHC-Fr, WB, IHC-FoFr, Flow Cyt (Intra), IHC-P
- Reacts with: Human, Rhesus monkey, African green monkey
- Isotype: IgG1
Related conjugates and formulations
Overview
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Product name
Anti-LAMP2 antibody [H4B4] - Lysosome Marker
See all LAMP2 primary antibodies -
Description
Mouse monoclonal [H4B4] to LAMP2 - Lysosome Marker -
Host species
Mouse -
Tested applications
Suitable for: ICC, ICC/IF, IHC-Fr, WB, IHC-FoFr, Flow Cyt (Intra), IHC-Pmore details -
Species reactivity
Reacts with: Human, Rhesus monkey, African green monkey
Does not react with: Mouse, Rat -
Immunogen
Tissue, cells or virus corresponding to Human LAMP2. Human adherent peripheral blood cells
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Positive control
- Flow Cyt (Intra): THP1 cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 8.20
Constituent: 100% Borate buffered saline -
Concentration information loading...
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Purity
Affinity purified -
Clonality
Monoclonal -
Clone number
H4B4 -
Isotype
IgG1 -
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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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab25631 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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ICC | (1) |
Use at an assay dependent concentration.
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ICC/IF | (11) |
Use at an assay dependent concentration.
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IHC-Fr | (1) |
Use at an assay dependent concentration.
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WB | (5) |
Use at an assay dependent concentration.
In our hands milk blocking is gives superior results to BSA blocking for this product. |
IHC-FoFr |
Use at an assay dependent concentration.
(PMID 19837699). |
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Flow Cyt (Intra) |
Use 0.5-1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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IHC-P | (1) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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ICC
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IHC-Fr
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. In our hands milk blocking is gives superior results to BSA blocking for this product. |
IHC-FoFr
Use at an assay dependent concentration. (PMID 19837699). |
Flow Cyt (Intra)
Use 0.5-1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, 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
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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 [H4B4] - Lysosome Marker (ab25631) at 1/500 dilution
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : Human liver lysate
Lane 4 : Human liver membrane fraction lysate
Lane 5 : Human skeletal muscle lysate
Lane 6 : Human brain lysate
Lane 7 : Human kidney lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
Performed under reducing conditions.
Additional bands at: 100 kDa (possible post-translational modification), 110 kDa (possible post-translational modification), 120 kDa (possible post-translational modification)
Exposure time: 20 minutesThis blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system under denaturing, reducing conditions. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. After transfer, the membrane was blocked for an hour in 3% milk before being incubated overnight at 4°C with mouse monoclonal [H4B4] to LAMP2 (ab25631; diluted 1:5000). Antibody binding was detected using peroxidase labelled goat anti-mouse IgG (ab97040; diluted 1:50000) for an hour at room temperature and visualised using ECL development solution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)Ab25631 staining human normal placenta. Staining is localized to the cytoplasm.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required. -
Overlay histogram showing THP1 cells stained with ab25631 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab25631, 0.5µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)This image is of an Abreciew submitted by Daniel Rodriguez.All lanes : Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631) at 1/500 dilution
Lane 1 : HEK293 cell lysate at 30 µg
Lane 2 : HepG2 at 30 µg
Lane 3 : THP-1 at 30 µg
Lane 4 : 3T3-L1 at 30 µl
Lane 5 : Mouse hepatocytes at 30 µl
Lane 6 : Rat hepatocytes at 30 µl
Secondary
All lanes : HRP conjugated Goat anti-Mouse IgG at 1/2000 dilution
Developed using the ECL technique.
Exposure time: 30 secondsAlong with THP-1 macrophages, other human cell lines were loaded, and the 110 kDa mature band for LAMP2 was detected in all the samples. On the other hand, mouse and rat cells were negative. The antibody works really good on human samples, detecting a single 110 kDa band but it's not suitable to use for mouse or rat samples.
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Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)This image was kindly supplied by Daniel Rodriguez by AbreviewAnti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631) at 1/500 dilution + whole cell lysate prepared from THP-1 macrophages at 30 µg
Secondary
Goat anti-mouse IgG conjugated to HRP at 1/2000 dilution
Developed using the ECL technique.
Observed band size: 110 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
Primary antibody incubated for 12 hours at 4°C.Gel running conditions: 12%Blocked with 5% milk for 1 hour at 25°C. -
Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (ab25631)This image is courtesy of an anonymous Abreview
Western blot analysis of Rhesus monkey primary retinal pigmented epthelium whole cell lysate and HeLa cell whole cell lysate (20µg/lane) labelling LAMP2 with ab25631 at 1/1000. An alkaline phosphatase-conjugated rabbit anti-mouse IgG was used as the secondary antibody.
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Intracellular Flow Cytometry analysis of Human T lymphocyte cell line Jurkat labeling LAMP2 with ab25631 at 1 µg/106 cells dilution (purple). A Goat Anti-Mouse IgG1, Human ads-FITC was used as the secondary antibody. Grey - Isotype Control, Mouse IgG1-UNLB, followed by Goat Anti-Mouse IgG1, Human ads-FITC.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (175)
ab25631 has been referenced in 175 publications.
- Palomba NP et al. Common and Rare Variants in TMEM175 Gene Concur to the Pathogenesis of Parkinson's Disease in Italian Patients. Mol Neurobiol 60:2150-2173 (2023). PubMed: 36609826
- Krishnan B et al. Active RB causes visible changes in nuclear organization. J Cell Biol 221:N/A (2022). PubMed: 35019938
- Wang S et al. Activation of Transcription Factor EB Alleviates Tubular Epithelial Cell Injury via Restoring Lysosomal Homeostasis in Diabetic Nephropathy. Oxid Med Cell Longev 2022:2812493 (2022). PubMed: 35082964
- Kumar R et al. Combinatorial Polycation Synthesis and Causal Machine Learning Reveal Divergent Polymer Design Rules for Effective pDNA and Ribonucleoprotein Delivery. JACS Au 2:428-442 (2022). PubMed: 35252992
- Fukushima S et al. Recruitment of LC3 by Campylobacter jejuni to Bacterial Invasion Site on Host Cells via the Rac1-Mediated Signaling Pathway. Front Cell Infect Microbiol 12:829682 (2022). PubMed: 35310852