Revision 1

#68534Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IF-F, IF-IC

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

65

Source/Isotype:

Rat IgG2a

UniProt ID:

#Q16625

Entrez-Gene Id:

100506658

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Frozen) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800

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.

Specificity / Sensitivity

Occludin (6B8A3) Rat mAb recognizes endogenous levels of total occludin protein. In some human tissue lysates, this antibody detects a band of unknown origin at approximately 95 kDa by western blot.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of mouse occludin protein.

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 the 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). Occludin is thought to be important in the assembly and maintenance of tight junctions. Differential phosphorylation of occludin at various residues may regulate its interaction with other tight junction proteins such as ZO-1 (3). VEGF-induced phosphorylation of occludin regulates tight junction stability and vascular permeability (4). Expression of occludin as well as claudin1 is required for infection of liver cells by hepatitis C virus (HCV) (5).

  1. Shin, K. et al. (2006) Annu Rev Cell Dev Biol 22, 207-35.
  2. Oliveira, S.S. and Morgado-Díaz, J.A. (2007) Cell Mol Life Sci 64, 17-28.
  3. Rao, R. (2009) Ann N Y Acad Sci 1165, 62-8.
  4. Murakami, T. et al. (2009) J Biol Chem 284, 21036-46.
  5. Ploss, A. et al. (2009) Nature 457, 882-6.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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.

Revision 1
#68534

Occludin (6B8A3) Rat mAb

Western Blotting Image 1: Occludin (6B8A3) Rat mAb Expand Image
Western blot analysis of extracts from various cell lines using Occludin (6B8A3) Rat mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of occludin protein in HeLa and Neuro-2a cells is consistent with the predicted expression pattern.
Western Blotting Image 2: Occludin (6B8A3) Rat mAb Expand Image
Western blot analysis of extracts from control DLD-1 cells (lane 1) or CRISPR/Cas9 occludin knockout (KO) DLD-1 cells (lane 2) using Occludin (6B8A3) Rat mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunofluorescence Image 1: Occludin (6B8A3) Rat mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse colon, labeled with Occludin (6B8A3) Rat mAb (left, green) and co-labeled with α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #76113 (right, red) and DAPI #4083 (right, blue).
Immunofluorescence Image 2: Occludin (6B8A3) Rat mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse kidney, labeled with Occludin (6B8A3) Rat mAb (left, green) and co-labeled with LAMP1 (E5N9Z) Rabbit mAb (Alexa Fluor® 594 Conjugate) #73294 (right, red) and DAPI #4083 (right, blue).
Immunofluorescence Image 3: Occludin (6B8A3) Rat mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse small intestine, labeled with Occludin (6B8A3) Rat mAb (left, green) and co-labeled with LAMP1 (E5N9Z) Rabbit mAb (Alexa Fluor® 594 Conjugate) #73294 (right, red) and α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #76113 (right, blue).
Immunofluorescence Image 1: Occludin (6B8A3) Rat mAb Expand Image
Confocal immunofluorescent analysis of control DLD-1 cells (left, positive) or CRISPR/Cas9 occludin knockout (KO) DLD-1 cells (right, negative) using Occludin (6B8A3) Rat mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).