FKBP51/FKBP5 Antibody (Hi51B)

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] -

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] - FKBP51/Hsp90 bind to GluR1-type AMPA receptors to regulate trafficking. A, Representative Western blottings form biotinylation assays of receptor endocytosis was performed on ex vivo slices, as described in Materials & Methods, rTgFKBP5 (N = 4; n = 8), WT (N = 2; n = 8), & tTA (N = 2; n = 8). Following labeling with Sulfo-NHS-SS biotin & chemical LTD (20 µM NMDA; 5 min) treatment, receptors were permitted to externalize at 30°C for the indicated times. B, The quantification ± SEM of multiple acquisitions is shown for GluR1. C, Representative Western blottings from anti-GluR1 co-immunoprecipitations & corresponding inputs from control & rTgFKBP5 mice immunoblotted as indicated. rTgFKBP5 (N = 2), WT (N = 2), & tTA (N = 2) total from two independent experiments. D, Representative Western blottings of anti-GluR1 co-immunoprecipitations & corresponding inputs from HEK293T cells transfected with GluR1 & FKBP51 or empty vector (EV) for 48 h were immunoblotted with antibodies as indicated. Just before harvest, cells were treated with 100 µM AMPA or PBS for 10 min to induce GluR1 receptor internalization. *p = 0.0286 by t-test of this time point. **p < 0.001 by two-way ANOVA. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30963102), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] -

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] - Detailed schematic & validation of the FKBP5 TRE transgene. A, To allow for site-directed, single copy insertion into the mouse genome in chromosome 11, the transgenic construct contained flanking attB sites via a PhiC31 integrase. The downstream Mp1 poly A tail will help maintain stable expression. To drive high expression, the transgenic construct included a tetracycline-response element (TRE) promoter made of seven repeats of the tetracycline operators used to drive high expression of the singly inserted FKBP5 gene in the presence of the tTA, & a weak minimal CMV promoter which produces low basal expression. B, Western blotting from HEK293T cells transfected with increasing amounts of FKBP5 TRE plasmid, as indicated, for 48 h. C, HEK293T cells were transfected with the indicated amounts of FKBP5 TRE & tTA plasmid, to ensure the tTA would drive high FKBP51 expression. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30963102), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] -

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] - FKBP51 expression & distribution in rTgFKBP5 mice. A, Expression of human & mouse FKBP5 in rTgFKBP5 mice expressed as fold change ± SEM compared to WT mice using qPCR; ***p < 0.001 by t test (N = 10) with three technical replicates. B, Western blotting showing FKBP51 levels in the hippocampus from rTgFKBP5, WT, FKBP5, & tTA mice. C, Western blotting showing levels of FKBP51 levels in the rTgFKBP5 hippocampus from 1 to 10 µg of protein loaded compared to 50 µg of protein from WT or FKBP51 mice. GAPDH levels are shown to confirm protein load. See Extended Data Figure 2-1 for more information on the antibody. D, 20× images of anti-FKBP51 staining from rTgFKBP5 mice. The entorhinal cortex (ECX), anterior cortex (ACX), CA1, CA3, & dentate gyrus (DG) are labeled. E, 20× images of anti-FKBP51 staining from rTgFKBP5 mice in the CA1, CA3, DG, ECX, & ACX. Scale bar = 100 µm; 10 µm (inset). F, Western blotting showing FKBP51 levels in the hippocampus, striatum, ACX, posterior cortex (PCX), interbrain, thalamus & hypothalamus, & cerebellum of a rTgFKBP5 mouse. G, Quantitation of FKBP51 proteins levels throughout the hippocampus (HPC), striatum (STR), ACX, PCX, interbrain, thalamus & hypothalamus (INTER), & cerebellum (CER), of rTgFKBP5 mice from multiple exposures. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30963102), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] -

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] - FKBP51/Hsp90 bind to GluR1-type AMPA receptors to regulate trafficking. A, Representative Western blottings form biotinylation assays of receptor endocytosis was performed on ex vivo slices, as described in Materials & Methods, rTgFKBP5 (N = 4; n = 8), WT (N = 2; n = 8), & tTA (N = 2; n = 8). Following labeling with Sulfo-NHS-SS biotin & chemical LTD (20 µM NMDA; 5 min) treatment, receptors were permitted to externalize at 30°C for the indicated times. B, The quantification ± SEM of multiple acquisitions is shown for GluR1. C, Representative Western blottings from anti-GluR1 co-immunoprecipitations & corresponding inputs from control & rTgFKBP5 mice immunoblotted as indicated. rTgFKBP5 (N = 2), WT (N = 2), & tTA (N = 2) total from two independent experiments. D, Representative Western blottings of anti-GluR1 co-immunoprecipitations & corresponding inputs from HEK293T cells transfected with GluR1 & FKBP51 or empty vector (EV) for 48 h were immunoblotted with antibodies as indicated. Just before harvest, cells were treated with 100 µM AMPA or PBS for 10 min to induce GluR1 receptor internalization. *p = 0.0286 by t-test of this time point. **p < 0.001 by two-way ANOVA. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30963102), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] -

Western Blot: FKBP51/FKBP5 Antibody (Hi51B) [NB110-96873] - Detailed schematic & validation of the FKBP5 TRE transgene. A, To allow for site-directed, single copy insertion into the mouse genome in chromosome 11, the transgenic construct contained flanking attB sites via a PhiC31 integrase. The downstream Mp1 poly A tail will help maintain stable expression. To drive high expression, the transgenic construct included a tetracycline-response element (TRE) promoter made of seven repeats of the tetracycline operators used to drive high expression of the singly inserted FKBP5 gene in the presence of the tTA, & a weak minimal CMV promoter which produces low basal expression. B, Western blotting from HEK293T cells transfected with increasing amounts of FKBP5 TRE plasmid, as indicated, for 48 h. C, HEK293T cells were transfected with the indicated amounts of FKBP5 TRE & tTA plasmid, to ensure the tTA would drive high FKBP51 expression. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30963102), licensed under a CC-BY license. Not internally tested by Novus Biologicals.