34-5400

Ikari A, Tonegawa C, Sanada A, Kimura T, Sakai H, Hayashi H, Hasegawa H, Yamaguchi M, Yamazaki Y, Endo S, Matsunaga T, Sugatani J
The Journal of biological chemistry 2014 May 9;289(19):13112-23

Enhanced immunohistochemical resolution of claudin proteins in glycolmethacrylate-embedded tissue biopsies.

Collins JE, Kirk A, Campbell SK, Mason J, Wilson SJ
Methods in molecular biology (Clifton, N.J.) 2011;762:371-82

Differential expression of claudin tight junction proteins in the human cortical nephron.

Kirk A, Campbell S, Bass P, Mason J, Collins J
Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association 2010 Jul;25(7):2107-19

Claudin-1 has tumor suppressive activity and is a direct target of RUNX3 in gastric epithelial cells.

Chang TL, Ito K, Ko TK, Liu Q, Salto-Tellez M, Yeoh KG, Fukamachi H, Ito Y
Gastroenterology 2010 Jan;138(1):255-65.e1-3

Endothelia of term human placentae display diminished expression of tight junction proteins during preeclampsia.

Liévano S, Alarcón L, Chávez-Munguía B, González-Mariscal L
Cell and tissue research 2006 Jun;324(3):433-48

Role of claudin interactions in airway tight junctional permeability.

Coyne CB, Gambling TM, Boucher RC, Carson JL, Johnson LG
American journal of physiology. Lung cellular and molecular physiology 2003 Nov;285(5):L1166-78

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1) and A549 (Lane 2). The blot was probed with Anti-Claudin-16 Rabbit Polyclonal Antibody (Product # 34-5400, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 34 kDa band corresponding to Claudin 16 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Claudin 16 was performed using 90% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Claudin 16 Rabbit Polyclonal Antibody (Product # 34-5400) at 2 µg/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) at 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 (Product # 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 junctional localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL