Claudin-15 : Turner Lab

Claudin-15 is a structural component of the tight junction that forms paracellular cation (sodium) channels.

Antibody validation This commercial rabbit antibody from ThermoFisher (38-9200) is an affinity isolated polyclonal antibody raised against a synthetic peptide derived from the C-terminal region of the mouse claudin-15 protein. Our validation included staining of formalin-fixed, paraffin-embedded human duodenum, where the antibody decorated tight junctions throughout the crypt-villus axis. The antibody also reacts with mice, and signal was similar in claudin-2 knockout mice (indicating that the antibody does not cross-react with claudin-2, which also forms a sodium channel). This antibody was characterized further in Holmes et al. (2006).

Immunostaining
Antigen retrieval with Target Retrieval Solution, pH9 (Agilent Technologies, Cat. #S2367) for 40 minutes at 95°C.
Primary and secondary incubations performed used the following reagents:

Antibody	Source	Catalog #	Lot #	RRID #	Working Dilution/Concentration
Rabbit anti-claudin-15 antibody	ThermoFisher	38-9200	QJ228299	AB_2533391	1.5 µg/ml
Mouse anti-E-cadherin antibody [M168]	Abcam	ab76055	GR216035	AB_1310159	1 µg/ml
Rat anti-actin antibody [MAC 237]	Abcam	ab50591	GR168034	AB_867488	8 µg/ml


Alexa Fluor 488 AffiniPure F(ab’)2 donkey anti-mouse IgG (H+L)	Jackson Immunoresearch	715-546-151	118237	AB_2341099	1.5 µg/ml
Alexa Fluor 594 AffiniPure F(ab’)2 donkey anti-rabbit IgG (H+L)	Jackson Immunoresearch	711-586-152	124065	AB_2340622	1.5 µg/ml
Alexa Fluor 647 AffiniPure F(ab’)2 donkey anti-rat IgG (H+L)	Jackson Immunoresearch	712-606-153	123491	AB_2340696	1.5 µg/ml
Hoechst 33342	Life Technologies	H3570			10 µg/ml