rabbit anti adam8  (Proteintech)

Journal: Journal of Healthcare Engineering

Article Title: ADAM8 Activates NLRP3 Inflammasome to Promote Cerebral Ischemia-Reperfusion Injury

doi: 10.1155/2021/3097432

Figure Lengend Snippet: Expression of ADAM8 in cerebral cortex of ischemia-reperfusion mice increased abnormally. (a) QPCR analysis of ADAM8 levels in mouse cerebral cortex, n = 3 mice per group. (b) Brain homogenates of the indicated group were obtained after reperfusion for 4/24/72 hours. And the level of the indicated proteins was analyzed with western blotting, n = 4 mice per group. All values are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. Sham group. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. MCAO group.

Article Snippet: Equal amounts of proteins (1 mg) were precleared with protein A + G-Sepharose beads for 1 hour at 4°C and were then incubated with 2 mg rabbit anti-ADAM8 antibodies (Abbexa, Cambridge, UK) and anti-ADAM8 antibodies (Cell Signaling Technology, Massachusetts, USA) overnight at 4°C.

Techniques: Expressing, Western Blot

Journal: Journal of Healthcare Engineering

Article Title: ADAM8 Activates NLRP3 Inflammasome to Promote Cerebral Ischemia-Reperfusion Injury

doi: 10.1155/2021/3097432

Figure Lengend Snippet: Strategy for generating the ADAM8 gene knockdown mice. (a) Effect of stereotactic cerebral injection, n = 4 mice per group. (b) Immunofluorescence analysis of adeno-associated virus in primary motor cortex region of mice 28 days after injection. Adeno-associated viruses carry green fluorescent protein (EGFP) and DAPI (blue) stains the nucleus (scale = 200 μ m). The magnification is 10x, n = 3 mice per group. (c) After reperfusion for 24 hours, the effects of different shRNA fragments were analyzed with western blotting, n = 5 mice per group. (d) The expression of ADAM8 in the cortex of mice in different treatment groups was detected by immunofluorescence staining, the magnification is 40x, and the scale bar was 50 μ m, n = 5 mice per group. All values are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. Sham Scramble shRNA group. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. MCAO Scramble shRNA group.

Article Snippet: Equal amounts of proteins (1 mg) were precleared with protein A + G-Sepharose beads for 1 hour at 4°C and were then incubated with 2 mg rabbit anti-ADAM8 antibodies (Abbexa, Cambridge, UK) and anti-ADAM8 antibodies (Cell Signaling Technology, Massachusetts, USA) overnight at 4°C.

Techniques: Knockdown, Injection, Immunofluorescence, Virus, shRNA, Western Blot, Expressing, Staining

Journal: Journal of Healthcare Engineering

Article Title: ADAM8 Activates NLRP3 Inflammasome to Promote Cerebral Ischemia-Reperfusion Injury

doi: 10.1155/2021/3097432

Figure Lengend Snippet: ADAM8 induced cerebral cortex injury induced in cerebral ischemia-reperfusion mice. (a) After reperfusion for 24 hours, neurological deficits of mice were assessed, n = 8 mice per group. (b) After reperfusion for 24 hours, brain sections of the indicated group were stained with 2,3,5-triphenyltetrazolium chloride (left) and analysis of infarct volume (right) was performed, n = 5 mice per group. (c) After reperfusion for 24 hours, cerebral blood flow of mice was recorded, n = 4 mice per group. (d) Nissl staining assays were performed in ischemic brain sections at 24 hours after I-R injury, n = 4 mice per group. All values are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. Sham Scramble shRNA group. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. MCAO Scramble shRNA group.

Article Snippet: Equal amounts of proteins (1 mg) were precleared with protein A + G-Sepharose beads for 1 hour at 4°C and were then incubated with 2 mg rabbit anti-ADAM8 antibodies (Abbexa, Cambridge, UK) and anti-ADAM8 antibodies (Cell Signaling Technology, Massachusetts, USA) overnight at 4°C.

Techniques: Staining, shRNA

Journal: Journal of Healthcare Engineering

Article Title: ADAM8 Activates NLRP3 Inflammasome to Promote Cerebral Ischemia-Reperfusion Injury

doi: 10.1155/2021/3097432

Figure Lengend Snippet: ADAM8 promoted the apoptosis of cerebral cortical neurons in cerebral ischemia and reperfusion mice. (a) Cleaved caspase-3 in the penumbra was detected by western blotting after reperfusion for 24 hours, n = 4 mice per group. (b) TUNEL and immunofluorescence analysis of the apoptosis cerebral in cortex penumbra at 24 hours after I-R injury. The magnification was 40x and the scale bar was 50 μ m, n = 3 mice per group. All values are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. Sham Scramble shRNA group. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. MCAO Scramble shRNA group.

Article Snippet: Equal amounts of proteins (1 mg) were precleared with protein A + G-Sepharose beads for 1 hour at 4°C and were then incubated with 2 mg rabbit anti-ADAM8 antibodies (Abbexa, Cambridge, UK) and anti-ADAM8 antibodies (Cell Signaling Technology, Massachusetts, USA) overnight at 4°C.

Techniques: Western Blot, TUNEL Assay, Immunofluorescence, shRNA

Journal: Journal of Healthcare Engineering

Article Title: ADAM8 Activates NLRP3 Inflammasome to Promote Cerebral Ischemia-Reperfusion Injury

doi: 10.1155/2021/3097432

Figure Lengend Snippet: ADAM8 promoted neuroinflammation in cerebral ischemia-reperfusion mice. (a) Western blot analysis of proteins was associated with inflammation after reperfusion for 24 hours, n = 3 mice per group. (b) and (c) immunofluorescence analysis of GFAP and Iba-1 in cortex penumbra at 24 hours after I-R. The magnification was 40x and the scale bar was 50 μ m, n = 3 mice per group. All values are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. Sham Scramble shRNA group. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. MCAO Scramble shRNA group.

Article Snippet: Equal amounts of proteins (1 mg) were precleared with protein A + G-Sepharose beads for 1 hour at 4°C and were then incubated with 2 mg rabbit anti-ADAM8 antibodies (Abbexa, Cambridge, UK) and anti-ADAM8 antibodies (Cell Signaling Technology, Massachusetts, USA) overnight at 4°C.

Techniques: Western Blot, Immunofluorescence, shRNA

Journal: Journal of Healthcare Engineering

Article Title: ADAM8 Activates NLRP3 Inflammasome to Promote Cerebral Ischemia-Reperfusion Injury

doi: 10.1155/2021/3097432

Figure Lengend Snippet: ADAM8 activated NLRP3 inflammasome in cerebral cortex of cerebral ischemia-reperfusion mice. (a) Western blotting reflected the relative level of indicated proteins in different groups after I-R 24 hours, n = 3 mice per group. (b) The interaction between ADAM8 and NLRP3 in cerebral cortex of I-R mice, n = 3 mice per group. (c) Immunofluorescence analysis of brain sections with antibodies specific for ADAM8 and NLRP3 (colocalization of ADAM8 with NLRP3). The magnification was 40x and the scale bar was 50 μ m. All values are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. Sham Scramble shRNA group. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. MCAO Scramble shRNA group.

Article Snippet: Equal amounts of proteins (1 mg) were precleared with protein A + G-Sepharose beads for 1 hour at 4°C and were then incubated with 2 mg rabbit anti-ADAM8 antibodies (Abbexa, Cambridge, UK) and anti-ADAM8 antibodies (Cell Signaling Technology, Massachusetts, USA) overnight at 4°C.

Techniques: Western Blot, Immunofluorescence, shRNA

Journal: Frontiers in Molecular Neuroscience

Article Title: ALWPs Improve Cognitive Function and Regulate Aβ Plaque and Tau Hyperphosphorylation in a Mouse Model of Alzheimer’s Disease

doi: 10.3389/fnmol.2019.00192

Figure Lengend Snippet: ALWPs significantly increase cell-surface levels of APP. (A) APP-H4 cells were treated with ALWPs (500 μg/ml) or PBS for 24 h, and cell-surface biotinylation assays were conducted with an anti-6E10 antibody. (B,C) Quantification of data from ( A ; surface APP and total APP; con, n = 8; ALWPs, n = 8). (D,E) Primary hippocampal neurons were transfected with GFP plasmid DNA for 24 h, followed by treatment with ALWPs (500 μg/ml) or PBS for 24 h and live cell-surface staining (con, n = 134 dendrites; ALWPs, n = 120 dendrites). (F) Primary hippocampal neurons were transfected with GFP plasmid DNA for 24 h, treated with ALWPs (500 μg/ml) or PBS for 24 h, and immunostained with an anti-ADAM17 antibody. (G) Quantification of data from ( F ; con, n = 67 dendrites; ALWPs, n = 71 dendrites). (H,J,L,N) Primary hippocampal neurons were treated with ALWPs (500 μg/ml) or PBS for 24 h and immunostained with anti-ADAM8, ADAM9, ADAM10, or ADAM12 antibodies. (I,K,M,O) Quantification of data from ( H,J,L,N ; con, n = 60 dendrites; ALWPs, n = 60 dendrites). * p < 0.05, *** p < 0.0001.

Article Snippet: We used the following primary antibodies for this study: mouse anti-4G8 (1:1,000, BioLegend, San Diego, CA, USA), mouse anti-6E10 (1:1,000, BioLegend, San Diego, CA, USA), rabbit anti-APP N-terminus (1:1,000, Sigma, Ronkonkoma, NY, USA), rabbit anti-ADAM8 (1:200, LSBio, Seattle, WA, USA), rabbit anti-ADAM9 (1:200, Abcam, UK), rabbit anti-ADAM10 (1:200, Abcam, UK), rabbit anti-ADAM12 (1:200, Abcam, UK), rabbit anti-ADAM17 (1:200, Abcam, UK), mouse anti-BACE1 (1:200, Abcam, UK), rabbit anti-neprilysin (1:200, Millipore, Burlington, MA, USA), mouse anti-AT100 (1:500, Invitrogen, Carlsbad, CA, USA), mouse anti-AT180 (1:500, Invitrogen, Carlsbad, CA, USA), and mouse anti-Tau5 (1:500, Invitrogen, Carlsbad, CA, USA).

Techniques: Transfection, Plasmid Preparation, Staining