Journal: Alzheimer's & dementia : the journal of the Alzheimer's Association

Article Title: Reduced synaptic proteins and SNARE complexes in Down syndrome with Alzheimer's disease and the Dp16 mouse Down syndrome model: Impact of APP gene dose.

doi: 10.1002/alz.12835

Figure Lengend Snippet: FIGURE 2 Reduction of SNARE complexes in AD-DS, AD, and Dp16 samples. Syntaxin 1A was present in a 35 kD monomeric form and in a 73 kD trimeric complex with SNAP25 and synaptobrevin 2 as well as other high molecular weight forms. A, D, G, Unboiled AD-DS (A), DS (D), and AD (G) SDS samples were loaded on SDS-PAGE and probed with the syntaxin 1A antibody. A boiled sample was also loaded as a control to demonstrate disruption of the complexes with heating. The arrow points to the 73 kD trimeric band. B, E, H, Profiles of relative intensities of the ladder of complexes from the representative samples marked with asterisks; the heaviest complexes are represented at the left end of the x-axis. C, F, I, Quantitation and statistical analysis of the levels of 73 kD trimeric SNARE complex in AD-DS and DS samples. Mann–Whitney test; *P < 0.05, **P < 0.01. J, Unboiled 2N, Dp16, and Dp16: App++−samples from mice aged 9 to 10 months were loaded on SDS-PAGE and probed with the syntaxin 1A antibody. Arrows point to the major 73 kD, 100–130 kD, and > 180 kD bands. K–M, Quantitation and statistical analysis of the levels of 73 kD, 100–130 kD and > 180 kD bands. One-way analysis of variance test followed by Newman–Keuls multiple comparison test. n = 3 mice for each category; *P < 0.05. C/, control for; AD-DS, Alzheimer’s disease with Down syndrome; DS, Down syndrome without Alzheimer’s disease; AD, Alzheimer’s disease; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis

Article Snippet: 3.1 Methods 3.1.1 Antibodies Mouse antibody anti-syntaxin 1A (1:5000; S0664; HPC-1) and rabbit antibody anti-APP (1:3000; A8717) were obtained from MilliporeSigma; rabbit antibody anti-SNAP25 (1:2000; 14903-1-AP), antiNPTX2 (10889-1-AP), mouse antibodies anti-synaptophysin (1:5000; 67864-1-Ig), anti-β-actin (1:10,000; 60008-1-Ig) were from Proteintech.

Techniques: High Molecular Weight, SDS Page, Control, Disruption, Quantitation Assay, MANN-WHITNEY, Comparison, Polyacrylamide Gel Electrophoresis

Journal: Alzheimer's & dementia : the journal of the Alzheimer's Association

Article Title: Reduced synaptic proteins and SNARE complexes in Down syndrome with Alzheimer's disease and the Dp16 mouse Down syndrome model: Impact of APP gene dose.

doi: 10.1002/alz.12835

Figure Lengend Snippet: FIGURE 4 Levels of selective biomarkers in the frontal cortex of AD-DS, DS, and the cortex of Dp16 mice. A, Western blotting analysis of the levels of NfL, NPTX2, and syntaxin1B in the frontal cortex of patients with DS, AD-DS, C/DS, or C/AD-DS. β-actin was used as a loading control. B, Quantitation and statistical analysis of the levels of NfL, NPTX2, and syntaxin 1B in female and male samples combined. Mann–Whitney test; *P < 0.05, **P < 0.01, ***P < 0.001. C, Western blotting analysis of the levels of NfL and NPTX2 in the cortex from 9- to 10-month-old and 20-month-old Dp16 mice. β-actin was used as a loading control. D, Quantitation and statistical analysis of the levels of NfL and NPTX2 in Dp16 mice. Unpaired Student t test; n = 4 mice for each category for 9- to 10-month-old mice, n = 3–4 mice for each category for 20-month-old mice; *P < 0.05. For all the proteins, the bands marked with (*) and ([) at the right molecular weight were quantitated as stated in the Methods section with the same policy applied to the bands for the same proteins from other samples. C/, control for; AD-DS, Alzheimer’s disease with Down syndrome; DS, Down syndrome without Alzheimer’s disease; AD, Alzheimer’s disease; NfL, neurofilament light; NPTX2, neuronal pentraxin 2

Article Snippet: 3.1 Methods 3.1.1 Antibodies Mouse antibody anti-syntaxin 1A (1:5000; S0664; HPC-1) and rabbit antibody anti-APP (1:3000; A8717) were obtained from MilliporeSigma; rabbit antibody anti-SNAP25 (1:2000; 14903-1-AP), antiNPTX2 (10889-1-AP), mouse antibodies anti-synaptophysin (1:5000; 67864-1-Ig), anti-β-actin (1:10,000; 60008-1-Ig) were from Proteintech.

Techniques: Western Blot, Control, Quantitation Assay, MANN-WHITNEY, Molecular Weight

Journal: F1000Research

Article Title: A guide to selecting high-performing antibodies for STXBP1 (UniProt ID: P61764 ) for use in western blot, immunoprecipitation, and immunofluorescence

doi: 10.12688/f1000research.160174.1

Figure Lengend Snippet: Lysates of U-87 MG WT and STXBP1 KO were prepared, and 40 μg of protein were processed for western blot with the indicated STXBP1 antibodies. The Ponceau stained transfers of each blot are presented to show equal loading of WT and KO lysates and protein transfer efficiency from the acrylamide gels to the nitrocellulose membrane. Antibody dilutions were chosen according to the recommendations of the antibody supplier. Antibody dilution used: ab109023** at 1/1000, ab124920** at 1/10 000, ab315893** at 1/1000, A5420 at 1/1000, NBP3-15097** at 1/2000, AF5675 at 1/200, 13414** at 1/1000, GTX114808 at 1/500, GTX114809 at 1/500, 67137-1-Ig* at 1/5000, MA5-35796** at 1/2000, PA1-742 at 1/500 (2 μg/ml). Predicted band size: 67.5 kDa. **Recombinant antibody, *Monoclonal antibody.

Article Snippet: Bio-Techne (Novus Biologicals) , NBP3-15097 , 230461 , AB_3094817 , recombinant mono , S05-4B6 , rabbit , n/a , Wb, IF.

Techniques: Western Blot, Staining, Membrane, Recombinant

Journal: F1000Research

Article Title: A guide to selecting high-performing antibodies for STXBP1 (UniProt ID: P61764 ) for use in western blot, immunoprecipitation, and immunofluorescence

doi: 10.12688/f1000research.160174.1

Figure Lengend Snippet: U-87 MG lysates were prepared, and immunoprecipitation was performed using 0.6 mg of lysate and 2.0 μg of the indicated STXBP1 antibodies pre-coupled to Dynabeads protein A or protein G. Samples were washed and processed for western blot with the indicated STXBP1 antibody. For western blot, NBP3-15097** was used at 1/2000. The Ponceau stained transfers of each blot are shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate, HC= antibody heavy chain. **Recombinant antibody, *Monoclonal antibody.

Article Snippet: Bio-Techne (Novus Biologicals) , NBP3-15097 , 230461 , AB_3094817 , recombinant mono , S05-4B6 , rabbit , n/a , Wb, IF.

Techniques: Immunoprecipitation, Western Blot, Staining, Recombinant

Journal: F1000Research

Article Title: A guide to selecting high-performing antibodies for STXBP1 (UniProt ID: P61764 ) for use in western blot, immunoprecipitation, and immunofluorescence

doi: 10.12688/f1000research.160174.1

Figure Lengend Snippet: U-87 MG WT and STXBP1 KO cells were labelled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio on coverslips. Cells were stained with the indicatedSTXBP1 antibodies and with the corresponding Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (WT), red (antibody staining) and far-red (KO) channels was performed. Representative images of the merged blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. When an antibody was recommended for immunofluorescence by the supplier, we tested it at the recommended dilution. The rest of the antibodies were tested at 1 and 2 μg/ml and the final concentration was selected based on the detection range of the microscope used and a quantitative analysis not shown here. Antibody dilution used: ab109023** at 1/100, ab124920** at 1/100, ab315893** at 1/500, A5420 at 1/50, AF5675 at 1/200, NBP3-15097** at 1/500, 13414** at 1/100, GTX114808 at 1/800, GTX114809 at 1/1000, 67137-1-Ig* at 1/2000, MA5-35796** at 1/100, PA1-742 at 1/1000. Bars = 10 μm. **Recombinant antibody, *Monoclonal antibody.

Article Snippet: Bio-Techne (Novus Biologicals) , NBP3-15097 , 230461 , AB_3094817 , recombinant mono , S05-4B6 , rabbit , n/a , Wb, IF.

Techniques: Staining, Immunofluorescence, Concentration Assay, Microscopy, Recombinant

Journal: Acta Neuropathologica

Article Title: Isoglutaminyl cyclase contributes to CCL2-driven neuroinflammation in Alzheimer’s disease

doi: 10.1007/s00401-015-1395-2

Figure Lengend Snippet: Cocktails of antibodies used for double and triple labeling immunohistochemistry

Article Snippet: isoQC 3285 syntaxin-6 , 1:100 1:100 , Rabbit Mouse , Probiodrug Abcam , Bio-donkey anti-rabbit + SA-Cy2 Cy3 donkey anti-mouse.

Techniques: Labeling