Anti chrebp antibody

Small intestine tissues were washed with formaldehyde (37°C, 1% final concentration) and then opened longitudinally on a glass plate. The mucosa was scraped and separated from the underlying muscle layers by a glass microscope slide. The scraped mucosal cells were then fixed in formaldehyde (37°C, 1% final concentration) for 10 min at room temperature. The chromatin immunoprecipitation (ChIP) assay was performed as described previously [28 (link), 29 (link)] using the EpiQuik Tissue Chromatin Immunoprecipitation kit (P-2003, EpiGentek, Farmingdale, NY). Chromatin was immunoprecipitated with control IgG or anti-ChREBP antibody (Novus Biologicals), purified, and then analyzed by qPCR using a CFX384 Touch^™ Real-Time PCR Detection System (Bio-Rad Laboratories, Inc.). The following primers were used: ChoRE1 forward 5′GATTTC CTG CCG CAT TCA GA3′, reverse 5′TTTTCAGAC CTC CCA GAT GGA3′; ChoRE2 forward 5′TCC ATC CAC ACA CTT TCA AAC C3′, reverse 5′CAA GCC ACG GCC AAC AG3′; ChoRE3 forward 5′TCC CCG GCT CAC CTC AA3′, reverse 5′TTC GGA GTG GGA GTC TGG TT3′; ChoRE4/5 forward 5′TGG TCA GTC CGG TAG CAG TTG3′, reverse 5′CCT TTG CAG GGC AGG CTA A3′; mNHE3 ChoRE1 forward 5′GGG AGG ATA TAG GGA ATT TG3′, reverse 5′CGA TAC TTG AAA CGT ATA TAT GT3′; mCyclophilin forward 5′GGT CTT TGG GAA GGT GAA AGA A3′, reverse 5′GCC ATT CCT GGA CCC AAA A3′.