Eif2ak3 Antibody

Immunohistochemistry staining of mouse mammary gland samples from lactating mice (L10) with Biorbyt's anti-PERK. Positive staining signal observed in wild type mouse sample with anti-PERK staining only (middle image), but not in the knock out mouse sample (right image) and pre-immune serum staining (left image) The anti-PERK was diluted 1:1000 in 5% goat serum in PBS and allowed to incubate for 2h at room temperature in a humidified chamber.

PERK-dependent miR-216b induction.(a) PERK+/+ and PERK−/− MEFs were treated with 500 nM TG for indicated times. MiR-216b was assessed by qPCR (left graph); PERK and CHOP were assessed by immunoblot (right). (b) MiR-216b levels were quantified by qPCR following exposure of cells to thapsigargin and a small-molecule PERK inhibitor (left). PERK, eIF2α-p and CHOP induction were assessed by immunoblot (right). (c–e) MEFs of the indicated genotype were treated with TG (500 nM) for indicated intervals. Protein extracts from these cells were immunoblotted for the proteins as indicated (lower panels) and miR-216b levels were quantified by qPCR (upper panels; n = 3). (f) CHOP−/− MEFs were transfected with vector or CHOP and 2 days later treated with TG (500 nM) for indicated intervals. Protein extracts from these cells were immunoblotted for CHOP and miR-216b levels quantified by qPCR (n = 3). (g) MiR-216b expression and (h) c-Jun mRNA levels were analysed in MMTV-Neu tumours from either PERK+/+ or a PERK−/− background. Data represent mean±s.d. of three independent observations. Statistical significance was analysed analysed by Student's t-test. (*P < 0.05, WT versus −/−).

PERK-dependent miR-216b induction.(a) PERK+/+ and PERK−/− MEFs were treated with 500 nM TG for indicated times. MiR-216b was assessed by qPCR (left graph); PERK and CHOP were assessed by immunoblot (right). (b) MiR-216b levels were quantified by qPCR following exposure of cells to thapsigargin and a small-molecule PERK inhibitor (left). PERK, eIF2α-p and CHOP induction were assessed by immunoblot (right). (c–e) MEFs of the indicated genotype were treated with TG (500 nM) for indicated intervals. Protein extracts from these cells were immunoblotted for the proteins as indicated (lower panels) and miR-216b levels were quantified by qPCR (upper panels; n = 3). (f) CHOP−/− MEFs were transfected with vector or CHOP and 2 days later treated with TG (500 nM) for indicated intervals. Protein extracts from these cells were immunoblotted for CHOP and miR-216b levels quantified by qPCR (n = 3). (g) MiR-216b expression and (h) c-Jun mRNA levels were analysed in MMTV-Neu tumours from either PERK+/+ or a PERK−/− background. Data represent mean±s.d. of three independent observations. Statistical significance was analysed analysed by Student's t-test. (*P < 0.05, WT versus −/−).

PERK-dependent miR-216b induction.(a) PERK+/+ and PERK−/− MEFs were treated with 500 nM TG for indicated times. MiR-216b was assessed by qPCR (left graph); PERK and CHOP were assessed by immunoblot (right). (b) MiR-216b levels were quantified by qPCR following exposure of cells to thapsigargin and a small-molecule PERK inhibitor (left). PERK, eIF2α-p and CHOP induction were assessed by immunoblot (right). (c–e) MEFs of the indicated genotype were treated with TG (500 nM) for indicated intervals. Protein extracts from these cells were immunoblotted for the proteins as indicated (lower panels) and miR-216b levels were quantified by qPCR (upper panels; n = 3). (f) CHOP−/− MEFs were transfected with vector or CHOP and 2 days later treated with TG (500 nM) for indicated intervals. Protein extracts from these cells were immunoblotted for CHOP and miR-216b levels quantified by qPCR (n = 3). (g) MiR-216b expression and (h) c-Jun mRNA levels were analysed in MMTV-Neu tumours from either PERK+/+ or a PERK−/− background. Data represent mean±s.d. of three independent observations. Statistical significance was analysed analysed by Student's t-test. (*P < 0.05, WT versus −/−).

Western blot analysis using Biorbyt's anti-PERK to detect PERK in cell lysates. 300 µg PERK over-expressing 293T cell lysate (lanes 1 & 2), or 800 ug wild type (Lanes 3 & 4), and PERK knock out (lanes 5 & 6) MEF cell lysate were immunoprecipated with 15 µl anti-PERK, followed by western blotting with 1:1000 dilution of anti-PERK in 5% milk/TBST buffer. Lane 1, 293T cells over-expressing Myc-PERK wt, Lane 2, 293T cells over-expressing Myc-PERK K618A.