Anti-Thrombospondin/THBS1 Antibody Picoband®

Western blot analysis of Thrombospondin using anti-Thrombospondin antibody (A00667-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: mouse liver tissue lysates,
Lane 3: HELA whole Cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Thrombospondin antigen affinity purified polyclonal antibody (Catalog # A00667-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Thrombospondin at approximately 165KD, 180KD. The expected band size for Thrombospondin is at 130KD.

Sub-FS may increase TSP-1/TGF-β1, excitatory synapse and glutamate levels. A – C Gray bands and normalized gray values of TSP-1 and TGF-β1 relative to GAPDH at 3 days (n = 4/group). D – H Gray bands and normalized gray values of PSD-95, Synapsin I and VGLUT1 relative to GAPDH at 3 days ( n = 4/group). I , J Number of positive PSD-95 signals and representative immunohistochemical results of PSD-95 (green) in EC at 3 days (scale bar = 5 μm, n = 4/group). Blue, DAPI. K – N Immunohistochemical results of VGLUT1 (green) and CAMKII (red) in EC at 3 days (scale bar = 10 μm, n = 4/group). Blue, DAPI. O Content of glutamate ( n = 6/group) in EC and CA3 and ( P ) ratio of normalized power spectra intensity of beta waves ( n = 5/group) at 3 days. Mean ± SEMs were presented. * P < 0.05, ** P < 0.01, *** P < 0.001 vs control group (One-way ANOVA).
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Reduced TSP-1 expression decreases susceptibility and neuronal injury induced by sub-FS stimuli and alleviates neuronal damage induced by subsequent FS stimuli. A Experimental design. B Threshold dosages of PTZ at P14 ( n = 6/group). C Threshold body temperature for induction of febrile seizures at P9 ( n = 8/group). D , E Analysis and representative images of FJB staining at 3 days after 41°C stimulus (scale bar = 50 μm). F – H Gray bands and normalized gray values of cleaved caspase-3 and caspase-3 relative to GAPDH at 3 days after 41°C stimulus. I , J Representative images (scale bar = 50 μm) and number of positive signals of FJB staining at 3 days after FS stimuli. K – M Gray bands and quantitative analysis of cleaved caspase-3 and caspase-3 at 3 days after FS stimuli. D – M n = 4/group. Mean ± SEMs were presented. * P < 0.05, ** P < 0.01, ***P < 0.001 vs siRNA-NC group (Unpaired T-test).
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Reduced TSP-1 expression decreases excitatory synapse and glutamate levels. Gray bands and normalized gray values of TSP-1/TGF-β1 ( A – C ), PSD-95/synapsin I ( D – F ) and VGLUT1 ( G , H ) at 24 hours and 3 days ( n = 4/group). The immunohistochemistry results of PSD-95 (green, scale bar = 3 μm, I , J ), VGLUT1 (green, scale bar = 10 μm, K , L ) and CAMKII (red, scale bar = 10 μm, M , N ) in EC. I – N n = 4/group, Blue, DAPI. O Ratio of normalized power spectra intensity of beta waves ( n = 5/group) and ( P ) glutamate content ( n = 6/group). Mean ± SEMs were presented. * P < 0.05, ** P < 0.01, *** P < 0.001 vs siRNA-NC group (Unpaired T-test).
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LSKL administration decreases excitatory synapse and glutamate levels. Gray bands and normalized gray values of TSP-1/TGF-β1 ( A – C ), PSD-95/synapsin I ( D – F ) and VGLUT1 ( G , H ) at 24 hours and 3 days ( n = 4/group). I , J Number of positive PSD-95 signals and representative immunohistochemical results of PSD-95 (green) in EC at 3 days (scale bar = 3 μm, n = 4/group). K , L Immunohistochemistry results of VGLUT1 (green) in EC at 3 days (scale bar = 10 μm, n = 4/group). M , N Representative immunohistochemical results of CAMKII (red) in EC at 3 days (scale bar = 10 μm, n = 4/group). Blue, DAPI. O Ratio of normalized power spectra intensity of beta waves ( n = 5/group) and ( P ) content of glutamate ( n = 6/group) at 3 days. Mean ± SEMs were presented. * P < 0.05, ** P < 0.01, *** P < 0.001 vs saline groups (Unpaired T-test).
Index in PubMed under a CC BY license. PMID: 38346981