Product Specification Sheet Claudin-5 (CLDN5) Antibody

 Cat # CLDN5-A Rabbit anti-Mouse Claudin-5 (CLDN5) antibody, affinity purified SIZE: 100 ug

Claudins are transmembrane found at tight Form & Storage of Antibodies junctions that control the passage of ions and molecules across an epithelial sheet and the movement of proteins and lipids between apical and basolateral domains of epithelial Affinity pure IgG cells. They form trafficking and signaling platforms involved in  Solution the regulation of growth, proliferation, and differentiation of cells; and they serve as a regulatory barrier ('gate') that Concentration: 0.5 mg/ml Volume: 200 ul separates fluid compartments of different compositions (e.g., Supplied in PBS pH 7.4 + 0.1% BSA in the kidney, intestine, and brain). Claudin-19 plays a role in magnesium transport.  Lyophilized powder Claudin-5 is a that in humans is encoded by Reconstitute powder in 200 ul PBS to 0.5 mg/ml the CLDN5 . The CLDN5 (claudins) gene is mapped to human 22q11.21 and codes for a Storage: transmembrane protein, which is a component of tight junction. The protein is a part of blood-brain barrier. Short-term: 4oC for 1 month

Source of Antigen or Antibodies Long-term: at –200C or below in suitable aliquots after reconstitution for 1 year. Do not expose to multiple free/thaw Uniprot: O54942 cycles or store working, diluted solutions.

Host: Rabbit Recommended Usage

Clonality: Polyclonal Western Blotting: 0.5-2 ug/ml Theoretical band size: 23 kDa Purification: Ammonium sulfate followed by peptide affinity Observed Band size: ~20 kDa. purification IHC: 1-5 ug/ml. Antigen retrieval in pH 6 sodium citrate Immunogen: 18 amino acid synthetic peptide derived from buffer is recommended. Mouse Claudin-5 C terminal Cytoplasmic region conjugated to KLH. *Above concentrations are a suggestion and user’s must optimize assay based on their conditions. Antibody may Species Reactivity: Mouse, Rat, Human work in other applications such as Flow Cytometry, ICC, or IP. These methods have not been tested by ADI. Cross reactivity: Peptide region used as an immunogen *This product is for In vitro research use only. exhibits 94% homology with Rat, Human, Monkey, Cat, ______Horse, Dog, and Pig Claudin-5. 89% with Bovine and Related materials available from ADI Xenopus laevis and 83% with Chicken. Reactivity has only Catalog# Description been confirmed in-house with Mouse, Rat, and Human samples. CLDN191-A Rabbit Anti-Mouse/Human Claudin-19 (CLDN19) IgG (aff pure) Subcellular Location: Plasma membrane CLDN1-A Rabbit anti-Mouse Claudin-1 Alternative name: Brain endothelial cell clone 1 protein, (CLDN1) antibody, affinity purified Lung-specific membrane protein CLDN7-A Rabbit anti-Human Claudin-7 Recommended Secondary Antibody: Goat anti-Rabbit (CLDN7) antibody, affinity purified IgG-HRP (ADI cat#20320) CLDN5-A 180824AC Negative Control: Non-immune Rabbit IgG (ADI cat# 20009-1).

Alpha Diagnostic Intl Inc., 6203 Woodlake Center Dr, San Antonio, TX 78244, USA; Email: [email protected]  (800) 786-5777; (210) 561-9515; Fax: (210) 561-9544; Web Site: www.4adi.com

25 ug of a Mouse Lung lysate was heated for 5 minutes at 95oC then separated on an ‘Any Kd’ SDS-PAGE gel (Biorad). The gel was run for ~1 hour and 30 minutes at 100V and transferred to a 0.2 um nitrocellulose membrane using the low MW settings on a Transblot Turbo (Biorad). The blot was blocked for 1 hour at room temperature with 1% Fish plasma (Aquablock, EastCoastBio). CLDN5-A was diluted with TBST+0.1% BSA to 1 ug/ml and incubated overnight at 4oC. Blot was washed with TBS-T 3 times for 5 minutes each. Goat anti-rabbit IgG HRP (ADI cat#20320) diluted 1:10,000 (50 ng/ml) in TBST+0.1% BSA and incubated for 1 hour at room temperature. The blot was washed with TBS-T 3 times for 5 minutes each. The blot was incubated with ADI Femto ECL substrate (ADI cat#80210) for 5 minutes then imaged on a LI-COR C-DIGIT at high sensitivity settings. Specific band is observed at ~20 kDa.

Alpha Diagnostic Intl Inc., 6203 Woodlake Center Dr, San Antonio, TX 78244, USA; Email: [email protected]  (800) 786-5777; (210) 561-9515; Fax: (210) 561-9544; Web Site: www.4adi.com

4 um FFPE Human skin was heated for 20 minutes at 60oC. The slide was deparaffinized then blocked with 2.5% normal goat serum for 30 minutes at room temperature. CLDN5-A was diluted with PBST+1% BSA to 2 ug/ml and incubated for overnight at 4oC. The slide was washed 3 times with PBST and incubated with 3% Hydrogen Peroxide for 10 minutes. The slide was washed 2 times in PBST then incubated with Goat anti- rabbit HRP Polymer detection reagent at room temperature for 30 minutes. The slide was washed with PBST 3 times and incubated with DAB for 3 minutes. The slide wad briefly washed with distilled water and counterstained with Gill’s II Hematoxylin for 1 minute. The slide was washed in running tap water until the water ran clear and blued in 0.1% Sodium Bicarbonate pH 9.6.

Alpha Diagnostic Intl Inc., 6203 Woodlake Center Dr, San Antonio, TX 78244, USA; Email: [email protected]  (800) 786-5777; (210) 561-9515; Fax: (210) 561-9544; Web Site: www.4adi.com