Immunofluorescence analysis of Marveld2/ Tricellulin was performed using 90% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with MARVELD2 / Tricellulin Rabbit Polyclonal Antibody (488400) at 2 ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cell junctional localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant protein derived from the C-terminal region of the human Tricellulin protein|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2-4 ug/ml|
|Immunofluorescence (IF)||2-4 ug/ml|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
Tight junctions form an important barrier of paracellular transport in epithelial cells. Sealing of two adjacent cells at bicellular tight junctions, a point where three adjacent cells are in contact with each other. Tricellulin is the first protein identified that specifically concentrates in tricellular tight junctions. This protein has four membrane spanning domains, similarly to claudins. Tricellulin expression is high in epithelium-derived tissues, such as small intestine, kidney and lung. Functional evidence for the role of tricellulin in tight junction formation comes from siRNA studies, where suppression of its expression leads to compromised epithelial barrier and tight junction formation. Loss of function in tricellulin mutants missing all or most of a conserved region in the cytosolic domain which binds to the cytosolic scaffolding protein ZO-1, indicate that interaction with other known tight junction proteins plays an important role for the function of tricellulin.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
ILDR1 deficiency causes degeneration of cochlear outer hair cells and disrupts the structure of the organ of Corti: a mouse model for human DFNB42.
48-8400 was used in western blot to study the pathogenesis of hearing loss caused by ILDR mutations.
|Sang Q,Li W,Xu Y,Qu R,Xu Z,Feng R,Jin L,He L,Li H,Wang L||Biology open (4:411)||2015|