- 33-1511 - Invitrogen Antibodies OCLN antibody

Submitted references Comparative Analysis of Cell-Cell Contact Abundance in Ovarian Carcinoma Cells Cultured in Two- and Three-Dimensional In Vitro Models.

Kutova OM, Sencha LM, Pospelov AD, Dobrynina OE, Brilkina AA, Cherkasova EI, Balalaeva IV
Biology 2020 Dec 4;9(12)

Long-Term In Vitro Expansion of Epithelial Stem Cells Enabled by Pharmacological Inhibition of PAK1-ROCK-Myosin II and TGF-β Signaling.

Zhang C, Lee HJ, Shrivastava A, Wang R, McQuiston TJ, Challberg SS, Pollok BA, Wang T
Cell reports 2018 Oct 16;25(3):598-610.e5

Dual action of highbush blueberry proanthocyanidins on Aggregatibacter actinomycetemcomitans and the host inflammatory response.

Ben Lagha A, LeBel G, Grenier D
BMC complementary and alternative medicine 2018 Jan 10;18(1):10

TRPV4 Regulates Tight Junctions and Affects Differentiation in a Cell Culture Model of the Corneal Epithelium.

Martínez-Rendón J, Sánchez-Guzmán E, Rueda A, González J, Gulias-Cañizo R, Aquino-Jarquín G, Castro-Muñozledo F, García-Villegas R
Journal of cellular physiology 2017 Jul;232(7):1794-1807

The impact of lactoferrin with different levels of metal saturation on the intestinal epithelial barrier function and mucosal inflammation.

Majka G, Więcek G, Śróttek M, Śpiewak K, Brindell M, Koziel J, Marcinkiewicz J, Strus M
Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine 2016 Dec;29(6):1019-1033

Expression of claudins -2 and -4 and cingulin is coordinated with the start of stratification and differentiation in corneal epithelial cells: retinoic acid reversibly disrupts epithelial barrier.

Ortiz-Melo MT, Sánchez-Guzmán E, González-Robles A, Valdés J, Gómez-Flores E, Castro-Muñozledo F
Biology open 2013 Feb 15;2(2):132-43

Identification of PKCzetaII: an endogenous inhibitor of cell polarity.

Parkinson SJ, Le Good JA, Whelan RD, Whitehead P, Parker PJ
The EMBO journal 2004 Jan 14;23(1):77-88

An in vitro airway wall model of remodeling.

Choe MM, Sporn PH, Swartz MA
American journal of physiology. Lung cellular and molecular physiology 2003 Aug;285(2):L427-33

Role of megalin (gp330) in transcytosis of thyroglobulin by thyroid cells. A novel function in the control of thyroid hormone release.

Marinò M, Zheng G, Chiovato L, Pinchera A, Brown D, Andrews D, McCluskey RT
The Journal of biological chemistry 2000 Mar 10;275(10):7125-37

Role of megalin (gp330) in transcytosis of thyroglobulin by thyroid cells. A novel function in the control of thyroid hormone release.

Marinò M, Zheng G, Chiovato L, Pinchera A, Brown D, Andrews D, McCluskey RT
The Journal of biological chemistry 2000 Mar 10;275(10):7125-37

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Occludin (OC-3F10), FITC was performed using 70% confluent log phase Caco-2 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 Occludin Monoclonal Antibody (OC-3F10), FITC (Product # 33-1511) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear junctional and nuclear localization. Panel e shows the isotype control. Panel f shows no primary antibody control. The images were captured at 60X magnification.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Occludin Antibody, FITC conjugate (OC-3F10) was done on 90% confluent log phase CaCo2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Occludin Antibody, FITC conjugate (OC-3F10) (Product # 33-1511) at 1µg/mL in 1% BSA and incubated for 3 hours 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 594 Phalloidin (Product # A12381). Panel d is a merged image showing cell junctional localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Occludin was achieved by transfecting Caco-2 cells with Occludin specific siRNA (Silencer® select Product # s458015 and s531865). Immunofluorescence analysis was performed on untransfected Caco-2 cells (panels a, d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with Occludin specific siRNA (panel c,f). Cells were fixed, permeabilized, and labelled with Occludin Monoclonal Antibody (OC-3F10), FITC (Product # 33-1511, 5 µg/mL). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962) and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Reduction of specific junctional localization was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to Occludin. The images were captured at 60X magnification.
Conjugate: Green dye

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3. TJ components detected in cultured RCE1(5T5) corneal epithelial cells. This was performed either by: ( A-J ) immunostaining, ( K-M ) end point RT-PCR, or ( N ) Western blot. (A) We used alpha-catenin as an indicator of the presence of adhesion complexes in the cultured epithelia. (B,G) Ocln, (D) ZO-1, (E) cldn-1, (C,I) cldn-4, and (F,H) cgn were easily located in cell borders of one day post-confluent cultures. (G-I) are transverse sections showing that TJs (arrows) were located at the cell boundaries of suprabasal cells. In (G), dashed line indicates the boundaries between a suprabasal cell and the basal cell layer; in (H-J) the dashed line indicate the basal side of the epithelium. Nuclei were stained either with propidium iodide or TO-PRO(r)-3. In 7-day confluent epithelia we found (K) the expression of cldn-1, -2, -4, but not of cldn-3 and -7; (L) cgn, ocln, and (M) ZO-1. Primers for cldn-3 and -7 were verified in PCR reactions using mouse kidney cDNA (K, m-kidney) that led to amplification products of the expected size and sequence. PR-P0 was used as an internal marker that does not change during the differentiation process. The antibodies were only appropriate to immunodetect cldn-1 and -4 and cgn (N); lanes 2 and 3 correspond to the loading control for the experiment. PCR reaction in the presence (+) or the absence of cDNA (-) (negative control). Results correspond to six duplicated experiments. A-D: Bar = 20 mum; E,F: Bar = 40 mum; G: Bar = 8 mum, and H-J: Bar
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Expression level of analyzed proteins of tight junctions in SKOV-3 and SKOV-3.ip cells cultured in monolayer and 3D in vitro models. ( A , C ). The distributions of SKOV-3 cells (left plot) and SKOV-3.ip cells (right plot) according to fluorescence intensity detected after staining with occludin-specific and ZO-1-specific antibodies (red--monolayer culture, blue--spheroids, green--collagen hydrogel); ( B , D ) Expression levels of occludin and ZO-1 in monolayer and 3D models denoted as relative fluorescence values calculated as a ratio of mean fluorescence intensity of cells stained with specific antibodies to mean fluorescence intensity of cells stained with antibodies of isotypic control. mnlr , monolayer; sph , spheroids, clgn ; collagen hydrogel. ""*"" indicates significant difference in RF level from monolayer culture (ANOVA, Holm-Sidak's multiple comparisons test, p < 0.05).
Conjugate: Green dye

33-1511

Antibody data