CFTR (human disease) : Turner Lab

CFTR (cystic fibrosis transmembrane conductance regulator) is an apical membrane chloride channel. Mutations cause cystic fibrosis.

Antibody validation
This commercial rabbit antibody from Abcam (ab59394) is an affinity isolated polyclonal antibody raised against a synthetic non-phosphopeptide derived from human CFTR around the phosphorylation site of serine 737. Our validation included staining of formalin-fixed, paraffin-embedded human duodenum, where CFTR is detected within the apical membrane of crypt epithelial cells. Expression is markedly reduced in villous epithelium.

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-CFTR antibody	Abcam	ab59394	GR139238	AB_940993	10 µg/ml
Mouse anti-E Cadherin antibody [M168]	Abcam	ab76055	GR216035	AB_1310159	2.5 µg/ml
Rat anti-ZO-1	Turner lab	6B6E4		AB_2783858	5 µg/ml


Alexa Fluor 488 AffiniPure F(ab’)2 donkey anti-mouse IgG (H+L)	Jackson Immunoresearch	715-546-151	118237	AB_2341099	3 µg/ml
Alexa Fluor 594 AffiniPure F(ab’)2 donkey anti-rabbit IgG (H+L)	Jackson Immunoresearch	711-586-152	114063	AB_2340622	3 µg/ml
Alexa Fluor 647 AffiniPure F(ab’)2 donkey anti-rat IgG (H+L)	Jackson Immunoresearch	712-606-153	123491	AB_2340696	3 µg/ml
Hoechst 33342	Life Technologies	H3570			10 µg/ml