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Immunofluorescence analysis of Claudin-16 was performed using 90% confluent log phase HEK-293 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 ABfinity™ Claudin-16 (10HCLC) Recombinant Rabbit Oligoclonal Antibody (710220) at 2ug/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 (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||Dog, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 256-277 of human claudin-16|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||3-4 µg/ml|
* 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.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A.
ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Tight junction group of proteins are a cell-to-cell adhesion structure in epithelial cells that constitute the epithelial junctional complex with adherens junctions and desmosomes. Tight junction strands are mainly composed of claudins, occludin, and JAM. Claudin-16 belongs to the Claudin family of protein which is composed of 23 integral membrane proteins. Claudins are responsible for the formation of tight junction strands and are connected with the actin cytoskeleton mediated by ZO-1. Claudin 16, also known as Paracellin-1, is a renal tight junction protein required for paracellular magnesium resorption. It is localized in the intercellular junctions of the epithelial cells of the thick ascending limb of Henle's loop (a TAL marker) are used to localize this specific structure of nephrons in kidney sections. Mg2+ resorption occurs in TAL, and Claudin-16 is required for this paracellular function. Failure in renal 'handling' of magnesium can result in hypermagnesemia, whose major direct toxicity implications are cardiovascular as well as renal failure. Recent studies have shown that the lack of Claudin-16 proteins contributes to the dysfunction of paracellular renal transport systems.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
CLDN16; HOMG3; hypomagnesemia 3; hypomagnesemia 3, with hypercalciuria and nephrocalcinosis; paracellin-1; PCLN-1; PCLN1; with hypercalciuria and nephrocalcinosis
CLDN16; HOMG3; PCLN1