Revision 1

#8683

Store at -20C

Tight Junction Antibody Sampler Kit

1 Kit

(6 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
CD2AP Antibody213520 µl80 kDaRabbit
Claudin-1 (D5H1D) Rabbit Monoclonal Antibody1325520 µl20 kDaRabbit IgG
ZO-1 (D7D12) Rabbit Monoclonal Antibody819320 µl220 kDaRabbit IgG
ZO-2 Antibody284720 µl150 kDaRabbit
ZO-3 (D57G7) Rabbit Monoclonal Antibody370420 µl140 kDaRabbit IgG
Afadin (D1Y3Z) Rabbit Monoclonal Antibody1353120 µl205 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Tight Junction Antibody Sampler Kit provides an economical means to evaluate the presence of a number of proteins involved in tight junctions. The kit contains enough primary antibodies to perform two western blot experiments per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Tight junctions, or zona occludens, form a continuous barrier to fluids across the epithelium and endothelium. They function in regulation of paracellular permeability and in the maintenance of cell polarity, blocking the movement of transmembrane proteins between the apical and basolateral cell surfaces (reviewed in 1). Tight junctions are composed of claudin and occludin transmembrane proteins, which join the junctions to the cytoskeleton (1,2). The claudin family is composed of 23 integral membrane proteins, and their expression, which varies among tissue types, may determine both the strength and properties of the epithelial barrier (2,3). Zona occludens proteins ZO-1, -2, and -3 (also known as TJP1, 2, and 3) are peripheral membrane adaptor proteins that link junctional transmembrane proteins such as occludin and claudin to the actin cytoskeleton (reviewed in 4). ZO-1 and ZO-2 are required for tight junction formation and function (5,6). In subconfluent proliferating cells, ZO-1 and ZO-2 have been shown to colocalize to the nucleus and play a role in transcriptional regulation (7-9). Exogenous expression of the amino terminal portion of ZO-3 exerts a dominant negative effect that interferes with assembly of tight junctions and adherens junctions (10). ZO-1 has been shown to interact with afadin prior to the formation of tight junctions (11). Recent work has also shown that afadin is involved in controlling the directionality of cell movement when it is localized at the leading edge of moving cells (12,13). CD2AP is a scaffolding protein that is thought to link membrane proteins to the cytoskeleton (14-16). It plays a role in the formation of tight junctions in specialized cell types such as the slit diaphragm of the kidney glomerulus (17). CD2AP is also involved in the immunological synapse between CD2-expressing T cells and antigen presenting cells (18). Research studies have shown that interaction between CD2AP and other cytoskeletal proteins may regulate the endocytosis of EGFR (16).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from A431 and MCF7 cells using Claudin-1 (D5H1D) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 1: Tight Junction Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using Afadin (D1Y3Z) Rabbit mAb.
undefined Image 2: Tight Junction Antibody Sampler Kit
Western blot analysis of extracts of K562, A431 and COS cells, using CD2AP Antibody.
undefined Image 3: Tight Junction Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell types using ZO-2 Antibody.
undefined Image 4: Tight Junction Antibody Sampler Kit
Western blot analysis of extracts from MCF-7, HT-29 and HCT-15 cells using ZO-3 (D57G7) XP® Rabbit mAb.
undefined Image 5: Tight Junction Antibody Sampler Kit
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 6: Tight Junction Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using ZO-1 (D7D12) Rabbit mAb.
undefined Image 7: Tight Junction Antibody Sampler Kit
Immunoprecipitation of claudin-1 from A-431 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Claudin-1 (D5H1D) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Claudin-1 (D5H1D) Rabbit mAb.
undefined Image 8: Tight Junction Antibody Sampler Kit
Immunoprecipitation of afadin from A-431 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Afadin (D1Y3Z) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Afadin (D1Y3Z) Rabbit mAb.
undefined Image 9: Tight Junction Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of A431 cells using ZO-2 Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
undefined Image 10: Tight Junction Antibody Sampler Kit
Confocal immunofluorescent analysis of MCF-7 cells using ZO-3 (D57G7) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 11: Tight Junction Antibody Sampler Kit
Immunoprecipitation and western blot analysis of extracts from Hep G2 cells using ZO-1 (D7D12) Rabbit mAb. Lane 1 is 10% input.
undefined Image 12: Tight Junction Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Claudin-1 (D5H1D) XP® Rabbit mAb.
undefined Image 13: Tight Junction Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Claudin-1 (D5H1D) XP® Rabbit mAb.
undefined Image 14: Tight Junction Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded cell pellets, A-431 (left) and MCF7 (right), using Claudin-1 (D5H1D) XP® Rabbit mAb.
undefined Image 15: Tight Junction Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human skin using Claudin-1 (D5H1D) XP® Rabbit mAb.
undefined Image 16: Tight Junction Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.