Journal: iScience

Article Title: Immunogenicity of COVID-19 vaccines and their effect on HIV reservoir in older people with HIV

doi: 10.1016/j.isci.2023.107915

Figure Lengend Snippet:

Article Snippet: BLOTTO solution , BioShop , Cat#SKI400.500.

Techniques: Virus, Recombinant, Cell Culture, Sterility, Enzyme-linked Immunosorbent Assay, Polymer, Staining, Plasmid Preparation, Software, Cytometry

Journal: Immunotherapy Advances

Article Title: Potential human immunotherapeutics for plague

doi: 10.1093/immadv/ltab020

Figure Lengend Snippet: Limit of detection of V antigen by Mab 7.3. V antigen was coated to a microtitre plate in the dilution range 5 µg/ml to 85 pg/ml in triplicate (0.1 ml/well) and then probed with a fixed concentration (800 ng/ml) of biotinylated Mab 7.3. After washing in PBS/Tween 20 and blocking with 2% milk powder in PBS (Blotto), binding was detected with streptavidin peroxidase (Pierce High Sensitivity Streptavidin-HRP, 21132) at 1:10,000 in Blotto and developed with 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS; SeraCare, 5120-0041) and the signal was read at OD414. The LOD was 20 ng/ml.

Article Snippet: After washing in PBS/Tween 20 and blocking with 2% milk powder in PBS (Blotto), binding was detected with streptavidin peroxidase (Pierce High Sensitivity Streptavidin-HRP, 21132) at 1:10,000 in Blotto and developed with 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS; SeraCare, 5120-0041) and the signal was read at OD 414 .

Techniques: Concentration Assay, Blocking Assay, Binding Assay

Journal: Materials Today Bio

Article Title: Nanocarriers of shRNA-Runx2 directed to collagen IV as a nanotherapeutic system to target calcific aortic valve disease

doi: 10.1016/j.mtbio.2023.100620

Figure Lengend Snippet: (A) Collagen IV expression in the aortic valve. Representative fluorescence images of aortic root cryosections from diabetic/hyperlipidemic ApoE-deficient mice and C57BL/6 mice stained for collagen IV (red) and counterstained with DAPI (blue) to identify the nuclei; Scalebar: 200 ​μm. (B) Localization of fluorescently labeled lipopolyplexes either targeted to collagen IV (Cp-LPP/shCtr) or non-targeted (S-LPP/shCtr) in organs harvested from diabetic/hyperlipidemic ApoE-deficient mice injected with lipopolyplexes. Measurements were recorded 90 ​min after retroorbital administration employing an IVIS Spectrum imaging system Caliper 200, by detection of Rhodamine B fluorescence λex ​= ​535 ​nm and λem ​= ​620 ​nm. (C) Quantification of total radiant efficiency in heart, aorta, liver, and aortic root. (D) Representative images of Rhodamine B-labeled Cp-LPP/shCtr and S-LPP/shCtr lipopolyplexes (red) distribution in the aortic valve sections of diabetic/hyperlipidemic ApoE-deficient mice. Nuclei labeled with DAPI (blue); white arrows depict the accumulation of lipopolyplexes. Scale bar: 300 ​μm. (E) Biochemical parameters for liver (ALT, AST, ALP) and kidney function (BUN and creatinine) measured in the plasma of C57BL/6 control mice (injected with PBS) or mice treated with lipopolyplexes targeted to collagen IV (Cp-LPP/shCtr) or non-targeted (S-LPP/shCtr). Data are mean ​± ​S.D. (n ​= ​3 mice for each group).

Article Snippet: To visualize the collagen IV expression, the cryosections were washed with warm PBS and blocked in TBS Blotto A (cat. no. SC-2333, Santa Cruz) with 1% BSA, 0.2% Tween (cat. no. 85113, ThermoFisher Scientific) and 1% cold fish gelatin (cat. no. G7041, Sigma) for 1 h at room temperature, followed by the overnight incubation at 4 °C with primary anti-collagen IV antibody (cat. no. PA5-79058, ThermoFisher Scientific) at 1:250 dilution in PBS with 3% BSA.

Techniques: Expressing, Fluorescence, Staining, Labeling, Injection, Imaging, Clinical Proteomics, Control