Immunohistochemical analysis of paraffin-embedded normal human small intestine using Claudin-2 (E1H9O) Rabbit mAb (IHC Formulated).
Immunohistochemical analysis of paraffin-embedded normal human kidney using Claudin-2 (E1H9O) Rabbit mAb (IHC Formulated).
Immunohistochemical analysis of paraffin-embedded mouse kidney using Claudin-2 (E1H9O) Rabbit mAb (IHC Formulated).
Immunohistochemical analysis of paraffin-embedded SW1463 (left) and PANC-1 (right) cell pellets using Claudin-2 (E1H9O) Rabbit mAb (IHC Formulated).
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.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
|SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) #8114||SignalStain® Boost IHC Detection Reagent (AP, Rabbit) #18653|
|SignalStain® DAB Substrate Kit #8059||SignalStain® Vibrant Red Alkaline Phosphatase Substrate Kit #76713|
|SignalStain® Vivid Purple Peroxidase Substrate Kit #96632|
NOTE: Use of detection reagents other than those specified in this protocol may require further optimization of the primary antibody to account for the different sensitivities of the detection reagents.
posted February 2010
revised June 2020
Protocol Id: 283
Claudin-2 (E1H9O) Rabbit mAb (IHC Formulated) recognizes endogenous levels of total claudin-2 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp196 of human claudin-2 protein.
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-1 is expressed primarily in keratinocytes (4) and normal mammary epithelial cells, but is absent or reduced in breast carcinomas and breast cancer cell lines (5,6).
Claudin-2 is expressed primarily in the proximal tubule of the normal mammalian kidney, where it regulates transepithelial ion (e.g., Na+, Cl-) reabsorption (7). Increased expression of claudin-2 has been reported in some cancer cell lines (8), including A549 lung adenocarcinoma cells, where its nuclear distribution was positively associated with enhanced proliferation (9).
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