Myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) peptide (MEVGWYRSPFSRVVHLYRNGK) was generated by the Protein Core laboratory of the Blood Research Institute, BloodCenter of Wisconsin. The 2.4G2 hybridoma was obtained from the American Tissue Culture Collection. Anti-mouse CD4-APC-eFluor 780, CD25-Alexa Fluor 700, CD11b-PE, CD11b-biotin, IL-17-Alexa Fluor 647, CD11c-Biotin, B220-Biotin, CD8-Biotin, CD11b-biotin, Foxp3-PE and streptavidin-PE Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-mouse B220-PE-Texas Red, IFN-γ-PE, anti-active caspase 3-FITC and anti-human Ki67-FITC were purchased from BD Biosciences (San Diego, CA). Anti-mouse Ly-6C-APC and Ly-6G-APC-Cy7 were purchased from Biolegend (San Diego, CA). Anti-mouse/rat Neuropilin-1-APC was obtained from R&D Systems (Minneapolis, MN). Monoclonal antibodies SMI-32 (anti-nonphosphorylated neurofilament-H) and SMI-99 (anti-myelin basic protein (MBP)) were purchased from Covance (Emeryville, CA). Strepavidin Alexa 405 and goat anti-mouse Alexa 546 (H + L) were purchased from Life Sciences Advanced Technologies (St. Petersburg, FL). Anti-Biotin microbeads were purchased from MiltenyiBiotec (Auburn, CA).
Anti active caspase 3 fitc
Manufactured by BD
Sourced in United States
Anti-active caspase 3-FITC is a fluorescence-labeled antibody that specifically binds to the active form of caspase 3, a key enzyme involved in the apoptosis (programmed cell death) pathway. This antibody can be used to detect and quantify the presence of active caspase 3 in cells.
Automatically generated - may contain errors
Lab products found in correlation
Foxp3 pe, by Thermo Fisher Scientific (1 mentions)
Ly6g apc cy7, by BioLegend (1 mentions)
Streptavidin pe cy5, by Thermo Fisher Scientific (1 mentions)
Ifn γ pe, by BD (1 mentions)
Anti mouse ly6c apc, by BioLegend (1 mentions)
Cd11b pe, by Thermo Fisher Scientific (1 mentions)
B220 biotin, by Thermo Fisher Scientific (1 mentions)
Cd11c biotin, by Thermo Fisher Scientific (1 mentions)
Cd11b biotin, by Thermo Fisher Scientific (1 mentions)
Il 17 alexa fluor 647, by Thermo Fisher Scientific (1 mentions)
Cd8 biotin, by Thermo Fisher Scientific (1 mentions)
Smi 32, by Fortrea (1 mentions)
Smi99, by Fortrea (1 mentions)
Anti biotin microbead, by Miltenyi Biotec (1 mentions)
Live dead fixable aqua, by Thermo Fisher Scientific (1 mentions)
Cytoflex s, by Beckman Coulter (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
3 protocols using anti active caspase 3 fitc
1
Murine Model of Experimental Autoimmune Encephalomyelitis
2
Flow Cytometric Analysis of DNA Damage
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The upregulation of DNA damage markers as response to TOP1-inhibiting ADCs were analyzed by a flow cytometry–based readout. For this, 30,000 HER2-positive SKBR-3 cells were incubated for 24 hours to 72 hours with 0.5 or 5 μg/mL of ADCs. After the end of the incubation time, cells were stained with LIVE/DEAD Fixable Aqua (Thermo Fisher Scientific) followed by fixation and permeabilization using the BD Cytofix/Cytoperm Kit (BD Biosciences) according to the manufacturer's instructions. Intracellular staining of DNA damage markers was performed using anti-cleaved PARP (Asp214) PE, anti-H2AX (pSer139) AF647, and anti-active caspase 3 FITC (all BD Biosciences) or respective isotype controls. Cells were acquired on a CytoFLEX S flow cytometer (Beckman Coulter) and the percentage of positive cells was determined.
3
Apoptosis Evaluation of THP-1 Cells
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THP-1 cells (1 × 106 cells/mL) were seeded into a 24-well plate and
incubated at 37°C overnight. After that, the cells were incubated with TAIIA
and/or 3MA for 3, 6, and 24 hours. Cultured cells were centrifuged at 1200 rpm
at room temperature for 5 minutes followed by washing with ice-cold PBS 3 times.
Afterward, pelleted cells were stained with anti-active caspase-3-FITC (BD
Pharmingen, San Diego, CA, USA) and PI in 500 μL apoptosis binding buffer for
15 minutes and analyzed by flow cytometry using a FACSVerse instrument, and the
data were analyzed using FlowJo software.
incubated at 37°C overnight. After that, the cells were incubated with TAIIA
and/or 3MA for 3, 6, and 24 hours. Cultured cells were centrifuged at 1200 rpm
at room temperature for 5 minutes followed by washing with ice-cold PBS 3 times.
Afterward, pelleted cells were stained with anti-active caspase-3-FITC (BD
Pharmingen, San Diego, CA, USA) and PI in 500 μL apoptosis binding buffer for
15 minutes and analyzed by flow cytometry using a FACSVerse instrument, and the
data were analyzed using FlowJo software.
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