Revision 3

#49564Store 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, IP, IF-IC

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

23

Source/Isotype:

Rabbit IgG

UniProt ID:

#O00501

Entrez-Gene Id:

7122

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:6400

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

Claudin-5 (E8F3D) Rabbit mAb recognizes endogenous levels of total claudin-5 protein.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro197 of human claudin-5 protein.

Background

Tight junctions, or zonula 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. Tight junctions are composed of claudin and occludin 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. Alteration in claudin protein expression pattern is associated with several types of cancer (2,3). 

Claudin-5, highly expressed in brain and lung tissue, is a primary component of the tight junctions of blood-brain barrier endothelial cells. The blood-brain barrier regulates homeostatic and pathological conditions in the brain by limiting passage of material circulating to the brain. The blood-brain barrier can also impede the effectiveness of small molecule therapeutics (4). Claudin-5 is also thought to regulate tumor cell motility brain metastasis (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. Hewitt, K.J. et al. (2006) BMC Cancer 6, 186.
  4. Greene, C. et al. (2019) Fluids Barriers CNS 16, 3.
  5. Jia, W. et al. (2014) Mol Med Rep 9, 779-85.

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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation 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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 3
#49564

Claudin-5 (E8F3D) Rabbit mAb

Western Blotting Image 1: Claudin-5 (E8F3D) Rabbit mAb Expand Image
使用 Claudin-5 (E8F3D) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对各种细胞系和组织的提取物进行蛋白质印迹法分析。Hela 细胞中 claudin-5 蛋白的阴性表达与预测的表达模式一致。
Immunoprecipitation Image 1: Claudin-5 (E8F3D) Rabbit mAb Expand Image
对 SCLC-21H 细胞提取物中的 claudin-5 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Claudin-5 (E8F3D) Rabbit mAb。使用 Claudin-5 (E8F3D) Rabbit mAb 进行蛋白质印迹法分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunofluorescence Image 1: Claudin-5 (E8F3D) Rabbit mAb Expand Image
使用 Claudin-5 (E8F3D) Rabbit mAb(绿色)、β-Actin (8H10D10) Mouse mAb #3700(红色)和 DAPI #4083(蓝色)对 HUVE 细胞(左图,阳性)和 Hela 细胞(右图,阴性)进行共聚焦免疫荧光分析。