anti p rip1  (Proteintech)

Journal: Clinical and experimental hypertension (New York, N.Y. : 1993)

Article Title: Disruption of the caspase-1/IL-1β axis alleviates myocardial Ischemia/Reperfusion injury via improvement of mitochondrial homeostasis and reduction of Pyroptosis.

doi: 10.1080/10641963.2025.2506619

Figure Lengend Snippet: Figure 4. Caspase-1 silencing reduces the secretion of IL-1β and inhibits myocardial injury due to cardiomyocyte pyroptosis. A, Western blot of caspase-1 and IL-1β proteins in myocardial tissues of I/R mice treated with sh-caspase-1#1 or sh-caspase-1#2. B, IL-1β serum levels in I/R mice treated with sh-caspase-1 measured by ELISA. C, LDH level measurement in the serum of I/R mice treated with sh-caspase-1. D, evaluation of cardiac physical function in I/R mice treated with sh-caspase-1 by echocardiography. E, myocardial infarct size in I/R mice treated with sh-caspase-1 detected by TTC staining. F, HE staining of cardiomyocyte injury in I/R mice treated with sh-caspase-1. G, cardiomyocyte pyroptosis in I/R mice treated with sh-caspase-1 evaluated by CaV3 (live cardiomyocytes, green) and EBD (necrotic cells, red) double staining. H, Western blot of RIP1, p-RIP1, RIPK3, and p-RIPK3, MLKL, and p-MLKL proteins in myocardial tissues of I/R mice treated with sh-caspase-1. I, morphological changes of myocardial cells in ultrathin sections of myocardial tissue were observed by transmission electron microscopy. n = 10 mice for each treatment. * p < .05.

Article Snippet: After 5% skimmed milk powder blocking, the membrane was probed with primary rabbit antibodies to GAPDH (ab9485, Abcam), Caspase-1 (sc -392 736, Santa Cruz Biotechnology), IL-1β (ab234437, Abcam), RIP1 (ab300617, Abcam), p-RIP1 (53286, Cell Signaling Technology, Shanghai, China), RIPK3 (ab62344, Abcam), p-RIPK3 (ab195117, Abcam), MLKL (ab255747, Abcam), p-MLKL (ab196436, Abcam) and then with secondary antibody peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H + L) (#111035003, Jackson ImmunoResearch).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Double Staining, Transmission Assay, Electron Microscopy

Journal: Clinical and experimental hypertension (New York, N.Y. : 1993)

Article Title: Disruption of the caspase-1/IL-1β axis alleviates myocardial Ischemia/Reperfusion injury via improvement of mitochondrial homeostasis and reduction of Pyroptosis.

doi: 10.1080/10641963.2025.2506619

Figure Lengend Snippet: Figure 5. Caspase-1 silencing suppresses H/R-induced cardiomyocyte pyroptosis in vitro. A-B, Western blot of caspase-1 protein in primary cardiomyocytes after 30 min of hypoxia and reoxygenation at 4, 6 and 8 h. C, viability of H/R-induced cardiomyocytes treated with sh-caspase-1 measured by CCK-8 assay. D, LDH release in H/ R-induced cardiomyocytes treated with sh-caspase-1. E, Representative images of pyroptosis of H/R-induced cardiomyocytes treated with sh-caspase-1 measured by PI (red)/DAPI (blue) double staining. F, PI+/DAPI+ H/R-induced cardiomyocytes treated with sh-caspase-1. G, Western blot of RIP1, p-RIP1, RIPK3, and p-RIPK3, MLKL, and p-MLKL proteins in H/R-induced cardiomyocytes treated with sh-caspase-1. Cell experiments were repeated at least three times independently. * p < .05.

Article Snippet: After 5% skimmed milk powder blocking, the membrane was probed with primary rabbit antibodies to GAPDH (ab9485, Abcam), Caspase-1 (sc -392 736, Santa Cruz Biotechnology), IL-1β (ab234437, Abcam), RIP1 (ab300617, Abcam), p-RIP1 (53286, Cell Signaling Technology, Shanghai, China), RIPK3 (ab62344, Abcam), p-RIPK3 (ab195117, Abcam), MLKL (ab255747, Abcam), p-MLKL (ab196436, Abcam) and then with secondary antibody peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H + L) (#111035003, Jackson ImmunoResearch).

Techniques: In Vitro, Western Blot, CCK-8 Assay, Double Staining

Journal: Clinical and experimental hypertension (New York, N.Y. : 1993)

Article Title: Disruption of the caspase-1/IL-1β axis alleviates myocardial Ischemia/Reperfusion injury via improvement of mitochondrial homeostasis and reduction of Pyroptosis.

doi: 10.1080/10641963.2025.2506619

Figure Lengend Snippet: Figure 7. IL-1β participates in the cardiomyocyte pyroptosis due to mitochondrial dysfunction by caspase-1. A, immunofluorescence staining of the co-localization of caspase-1 and IL-1β in the H/R-induced cardiomyocytes. B, Western blot of IL-1β protein in H/R-induced cardiomyocytes treated with sh-caspase-1. C, IL-1β levels in the supernatant of H/R-induced cardiomyocytes treated with sh-caspase-1 measured by ELISA. D, viability of H/R-induced cardiomyocytes treated with sh-caspase-1 measured by CCK-8, E, LDH release in H/R-induced cardiomyocytes treated with sh-caspase-1. F, Representative images of pyroptosis of H/R-induced cardiomyocytes treated with sh-caspase-1 measured by PI (red)/DAPI (blue) double staining. G, PI+/DAPI+ H/R-induced cardiomyocytes treated with sh-caspase-1. H, Western blot of RIP1, p-RIP1, RIPK3, and p-RIPK3, MLKL, and p-MLKL proteins in H/R-induced cardiomyocytes treated with sh-caspase-1. Cell experiments were repeated at least three times independently. * p < .05.

Article Snippet: After 5% skimmed milk powder blocking, the membrane was probed with primary rabbit antibodies to GAPDH (ab9485, Abcam), Caspase-1 (sc -392 736, Santa Cruz Biotechnology), IL-1β (ab234437, Abcam), RIP1 (ab300617, Abcam), p-RIP1 (53286, Cell Signaling Technology, Shanghai, China), RIPK3 (ab62344, Abcam), p-RIPK3 (ab195117, Abcam), MLKL (ab255747, Abcam), p-MLKL (ab196436, Abcam) and then with secondary antibody peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H + L) (#111035003, Jackson ImmunoResearch).

Techniques: Immunofluorescence, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Double Staining

Journal: Journal of Animal Science and Biotechnology

Article Title: Necroptosis contributes to deoxynivalenol-induced liver injury and inflammation in weaned piglets

doi: 10.1186/s40104-024-01117-1

Figure Lengend Snippet: Effects of dietary or gavage with DON on protein expression of necroptosis signals in liver cells of piglets. A The protein expression of necroptosis signals after dietary DON exposure. B Representative bands of necroptosis signals after dietary DON exposure. C The protein expression of necroptosis signals after DON gavage. D Representative bands of necroptosis signals after DON gavage. Values are means ± SE, n = 6. a,b Values without a common letter differ significantly ( P < 0.05). DON, Deoxynivalenol; p-MLKL, Phosphorylated mixed lineage kinase domain-like protein; p-RIP1, Phosphorylated receptor interacting protein kinase 1; p-RIP3, Phosphorylated receptor interacting protein kinase 3; t-MLKL, Total mixed lineage kinase domain-like protein; t-RIP1, Total receptor interacting protein kinase 1; t-RIP3, Total receptor interacting protein kinase 3

Article Snippet: Specific primary antibodies including mouse anti-t-RIP1 (1:1,000, LifeSpan BioSciences, Seattle, Washington, USA), rabbit anti-phosphorylated RIP1 (p-RIP1) (1:2,000, Cell Signaling Technology, Boston, Massachusetts, USA), mouse anti-t-RIP3 (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-phosphorylated RIP3 (p-RIP3) (1:2,000, Cell Signaling Technology, Boston, Massachusetts, USA), rabbit anti-t-MLKL (1:1,000, Cell Signaling Technology, Boston, Massachusetts, USA), rabbit anti-phosphorylated MLKL (p-MLKL) (1:1,000, Cell Signaling Technology, Boston, Massachusetts, USA) and mouse anti-β-actin (1:10,000, Sigma Aldrich, St. Louis, Missouri, USA).

Techniques: Expressing

Journal: Journal of Animal Science and Biotechnology

Article Title: Necroptosis contributes to deoxynivalenol-induced liver injury and inflammation in weaned piglets

doi: 10.1186/s40104-024-01117-1

Figure Lengend Snippet: Effects of Nec-1 on protein expression of necroptosis signals in liver of piglets after DON gavage. Piglets were given a gavage with 2 mg/kg BW DON or an equal volume of normal saline after intraperitoneal injection of 0.5 mg/kg BW Nec-1 or an equal volume of 5% dimethylsulfoxide (DMSO). Pigs were euthanized at 6 h after DON or saline gavage. A–I Protein expression of necroptosis signals. J Representative bands. Values are means ± SE, n = 6. a,b Values without a common letter differ significantly ( P < 0.05). DON, Deoxynivalenol; p-MLKL, Phosphorylated mixed lineage kinase domain-like protein; p-RIP1, Phosphorylated receptor interacting protein kinase 1; p-RIP3, Phosphorylated receptor interacting protein kinase 3; t-MLKL, Total mixed lineage kinase-like protein; t-RIP1, Total receptor interacting protein kinase 1; t-RIP3, Total receptor interacting protein kinase 3

Article Snippet: Specific primary antibodies including mouse anti-t-RIP1 (1:1,000, LifeSpan BioSciences, Seattle, Washington, USA), rabbit anti-phosphorylated RIP1 (p-RIP1) (1:2,000, Cell Signaling Technology, Boston, Massachusetts, USA), mouse anti-t-RIP3 (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-phosphorylated RIP3 (p-RIP3) (1:2,000, Cell Signaling Technology, Boston, Massachusetts, USA), rabbit anti-t-MLKL (1:1,000, Cell Signaling Technology, Boston, Massachusetts, USA), rabbit anti-phosphorylated MLKL (p-MLKL) (1:1,000, Cell Signaling Technology, Boston, Massachusetts, USA) and mouse anti-β-actin (1:10,000, Sigma Aldrich, St. Louis, Missouri, USA).

Techniques: Expressing, Saline, Injection