primary polyclonal antibodies against claudin–5  (MyBiosource Biotechnology)

Journal: bioRxiv

Article Title: Neurovascular mitochondrial susceptibility impacts blood-brain barrier function and behavior

doi: 10.1101/2024.02.15.580544

Figure Lengend Snippet: ( A ) Representative plot of oxygen consumption rate by a bezafibrate-treated and untreated 22qDS iBBB line during the Seahorse XFe96 Analyzer mitochondrial stress test ( n = 4 22qDS lines, 7 replicates per condition). ( B ) Quantification of OXPHOS parameters for each 22qDS line comparing bezafibrate-treated and untreated conditions ( n = 4 22qDS lines, 7 replicates per condition, paired t test). ( C ) Schematic is describing key regulators of mitochondrial dynamics. ( D ) Quantification of gene transcripts for fusion and fission ( n = 4 22qDS lines, paired t test). ( E ) Representative time course of TEER fold change following bezafibrate treatment in a 22qDS iBBB line, normalized to untreated ( n = 6-11 replicates per condition, two-way ANOVA). ( F ) Quantification of TEER at 36 hours following bezafibrate treatment ( n = 4 22qDS lines, 3-4 replicates per condition – 2 experiments per line, paired t test). ( G ) Immunofluorescence of claudin-5 (green) in 22qDS in untreated (top) compared to bezafibrate treatment (bottom). Error bars ± SEM. Scale bar: 10 μm.

Article Snippet: Primary antibody rabbit polyclonal antibodies against claudin-5 (Life Technologies) was diluted 1:300 in 3% normal donkey serum and incubated overnight at 4°C.

Techniques: Immunofluorescence

Journal: bioRxiv

Article Title: Neurovascular mitochondrial susceptibility impacts blood-brain barrier function and behavior

doi: 10.1101/2024.02.15.580544

Figure Lengend Snippet: ( A ) Quantification of mitochondrial content by Mitotracker fluorescence ( n = 7). ( B ) Representative EM images and ( C ) quantification of intramitochondrial density in BBB-ECs from treated and untreated 22qMc. ( D ) Representative immunofluorescent staining hippocampal vessels for leakiness (IgG, red) and claudin-5 (green). ( E ) Quantification of BBB leakiness as measured by serum IgG extravasation and ( F ) quantification of claudin-5 in hippocampal vessels in treated and untreated 22qMc ( n = 7-8). Error bars ± SEM. Scale bar: *, p < 0.05, ** p < 0.01 by nested t test.

Article Snippet: Primary antibody rabbit polyclonal antibodies against claudin-5 (Life Technologies) was diluted 1:300 in 3% normal donkey serum and incubated overnight at 4°C.

Techniques: Fluorescence, Staining

Journal: Acta Pharmacologica Sinica

Article Title: Increased BBB permeability contributes to EGCG-caused cognitive function improvement in natural aging rats: pharmacokinetic and distribution analyses

doi: 10.1038/s41401-019-0243-7

Figure Lengend Snippet: The permeability of blood–brain barrier (BBB) was apparently increased in the aging with cognitive impairment (CI) groups. a The absorbance of EB staining was increased in the aging with CI groups (n = 6). b The ultra-structures of tight junctions (TJs) of BBB in the aging with CI groups were changed by transmission electron microscopy (TEM) scanning. c The expressions of the TJ-associated proteins, such as claudin-5, occludin, and ZO-1 were down-regulated in the aging with CI groups. **P < 0.01, compared with the control group

Article Snippet: Nonspecific binding sites on the membrane were blocked by PBS containing 0.1% Tween-20 (PBST) with 5% bovine serum albumin for 1 h, followed by incubation with rabbit polyclonal primary antibodies against claudin-5, occludin, ZO-1 (1:1000, Sigma, St. Louis, MO, USA), and β-actin (1:1000, Santa Cruz) overnight at 4 °C.

Techniques: Permeability, Staining, Transmission Assay, Electron Microscopy