Thymuses from untreated and SL-TBI treated mice were harvest, enzymatically digested and stained with cell surface markers for thymus populations. Cells were further stained for caspase 1 cleavage with Caspase-1 (active) Staining Kit (Abcam, ab219935), or fixed for caspase 3 analysis using Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor® 488 Conjugate) (Cell signaling, 9669S). Apoptosis and pyroptosis was assessed by adding Annexin V-FITC (Biolegend, 640906), Annexin V binding buffer (BioLegend, 422201) and Propidium Iodide (Invitrogen, BMS500PI). Gasdermin D was measured in freshly isolated thymocytes using Gasdermin D (mouse) ELISA Kit (Adipogen Life Sciences, AG-45B-0011-KI01). Lactate dehydrogenase was assessed from the supernatant of harvested thymocytes using Lactate Dehydrogenase assay (Abcam, ab102526).
Caspase 1 active staining kit
Manufactured by Abcam
The Caspase-1 active staining kit is a tool designed to detect and visualize active caspase-1 in cells. It provides a reliable method for studying the activation of this key enzyme involved in inflammatory processes.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Annexin 5 binding buffer, by BioLegend (1 mentions)
Annexin 5 fitc, by BioLegend (1 mentions)
Alexa fluor 488 conjugate, by Cell Signaling Technology (1 mentions)
Gasdermin d mouse elisa kit, by Adipogen (1 mentions)
Nigericin, by InvivoGen (1 mentions)
4 protocols using caspase 1 active staining kit
1
Thymic Apoptosis and Pyroptosis Evaluation
2
Inflammasome Activation and Regulation
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BMDMs were seeded as described above and treated with Pam3CSK4 for 24 h followed by fingolimod and febuxostat treatments for 3 h followed by nigericin (5 μM; Invivogen) for 3 h. ROS generation, PP2A activity and IL-1β levels were determined as described above. In a separate set of experiments, BMDMs (50,000 cells per well) were seeded in 96 well plates and stimulation and treatments were conducted as described above and caspase-1 activity was measured using Caspase 1 (active) Staining Kit (Abcam).
3
Caspase-1 Expression in Primed BMDMs
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To determine the expression of caspase-1, primed BMDMs were stimulated with 20 µg/ml CTB and 2 µg/ml LPS O111:B4 for 4 and 16 hr, respectively, washed twice with PBS, stained using a caspase-1 active staining kit (Abcam) including caspase-1 staining (fluorescent green) and DAPI staining (fluorescent blue) according to the manufacturer’s protocol and analyzed using a confocal microscope.
4
Caspase-1 Activity Assay in Primed BMDMs
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To determine the expression of Caspase-1, primed BMDMs were stimulated with 20µg/ml CTB and 2µg/ml LPS O111:B4 for 4 and 16 hours respectively, washed twice with PBS, stained using a caspase-1 active staining kit (Abcam) according to the manufacturer's protocol and analyzed using a confocal microscope.
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