Product nameAnti-LYVE1 antibody
See all LYVE1 primary antibodies
DescriptionRabbit polyclonal to LYVE1
Tested applicationsSuitable for: IHC-Fr, Flow Cyt, IHC-P, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Rat
- human colon carcinoma
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Concentration information loading...
PurityProtein A purified
Purification notesProtein-A Chromatography (+his tag depleted).
Primary antibody notesThe lymphatic vasculature forms a second circulatory system that drains extracellular fluid from the tissues and provides an exclusive environment in which immune cells can encounter and respond to foreign antigen. Recently a number of interesting molecules have been identified that may be exploited as markers for lymphatic endothelium, including the hyaluronan receptor LYVE1, PALE, VEGFR3, podoplanin.
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab10278 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use a concentration of 6 - 30 µg/ml.
Fix sections for 10 min at -20°C in MeOH.
|Flow Cyt||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 2 µg/ml.|
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 35-45 kDa (predicted molecular weight: 35 kDa).Can be blocked with Recombinant Human LYVE1 protein (ab54341).|
FunctionLigand-specific transporter trafficking between intracellular organelles (TGN) and the plasma membrane. Plays a role in autocrine regulation of cell growth mediated by growth regulators containing cell surface retention sequence binding (CRS). May act as a hyaluronan (HA) transporter, either mediating its uptake for catabolism within lymphatic endothelial cells themselves, or its transport into the lumen of afferent lymphatic vessels for subsequent re-uptake and degradation in lymph nodes.
Tissue specificityMainly expressed in endothelial cells lining lymphatic vessels.
Sequence similaritiesContains 1 Link domain.
Cellular localizationMembrane. Localized to the plasma membrane and in vesicles near extranuclear membranes which may represent trans-Golgi network (TGN) and endosomes/prelysosomeal compartments. Undergoes ligand-dependent internalization and recycling at the cell surface.
- Information by UniProt
- Cell surface retention sequence-binding protein 1 antibody
- CRSBP 1 antibody
- CRSBP-1 antibody
Immunohistochemistry (Frozen sections) analysis of human colon carcinoma tissue sections labelling LYVE1 with ab10278.
ab10278 (2µg/ml) staining LYVE1 in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is lymphatic endothelium staining of lymphatic ducts where blood vessel endothelium and smooth muscle is wholly negative.
Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
All lanes : Anti-LYVE1 antibody (ab10278) at 1 µg/ml
Lane 1 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 35 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Additional bands at: 22 kDa, 55 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutes
LYVE-1 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
Flow Cytometry analysis of human dermal microvascular endothelial cells (HDMVEC) labelling LYVE1 with ab10278.
Rat skin was fixed with paraformaldehyde in 15% saturated picric acid solution for 4hr. Prior to sectioning, the specimen was infiltrated in O.C.T. and frozen in isopentane. The frozen specimen was sectioned these were rinsed in PBS for 15 min to remove O.C.T. and incubated in a 3% sodium deoxycholate solution. The specimens were rinsed twice with distilled water and then with PBS three times. The sections were incubated in 10% normal goat serum for 12 hr at 4°C, then for 12 hr with ab10278. After washing with PBS, the specimens were incubated with Alexa Fluor® 555-conjugated goat anti-rabbit IgG (H+L) (1:500), for 12 hr at 4°C. The cell nuclei were counterstained with YoYo-1. Images were obtained by using confocal microscope.
This product has been referenced in:
- Nishida-Fukuda H et al. Ectodomain Shedding of Lymphatic Vessel Endothelial Hyaluronan Receptor 1 (LYVE-1) Is Induced by Vascular Endothelial Growth Factor A (VEGF-A). J Biol Chem 291:10490-500 (2016). Read more (PubMed: 26966180) »
- Klar AS et al. Analysis of blood and lymph vascularization patterns in tissue-engineered human dermo-epidermal skin analogs of different pigmentation. Pediatr Surg Int 30:223-31 (2014). Human . Read more (PubMed: 24363089) »