Cell Viability Assays for Neuroscience Research

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Figure 1. Fluorescent Microscopy Analysis of THP-1 Suspension Cells THP-1 cells were stained with Poly (active) Caspase FAM FLICA® Inhibitor Reagent (green), and Hoechst 33342 (Blue). Cells were incubated with staurosporine to induce apoptosis. Two photos were taken and superimposed. Only one cell of the three cells is apoptotic (middle) – it is stained positive for caspase activity with FAM-VAD-FMK. Data courtesy of Dr. Brian W. Lee.Figure 2. Caspase Activity in Jurkat Cells Jurkat cells (T lymphocytes) were labeled with our Green Fluorescent Poly (active) Caspase FAM FLICA® Inhibitor Reagent (FAM-VAD-FMK) (Cat. 92) and viewed under a fluorescence microscope. The grey DIC image (B) reveals five cells in the field of view, but only four of them fluoresce green (A). Four out of five cells are apoptotic and have active caspases present (green). Data courtesy of Dr. Brian W. Lee, ICT.
ImmunoChemistry Technologies Green Fluorescent Poly (active) Caspase FAM FLICA® Assay Kit (FAM-VAD-FMK)
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Figure 1. Jurkat cells were stimulated with Staurosporine for 3 hours (B) or DMSO (A). Mito Flow reagent was added according to the protocol, incubated for 30 minutes and analyzed by flow cytometry: Ex:488nm Em: FL2 channel.
ImmunoChemistry Technologies Mito Flow - Mitochondrial Membrane Potential Detection Kit
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Figure 1. Assay principleFigure 2. Detection of Hypochlorite ( -OCl ) in neutrophils. Neutrophils were isolated from porcine blood, washed in Krebs-Ringers phosphate buffer and seeded in glass bottom dishes. The cells were then loaded with APF (10μM final) by incubation for 30 minutes at room temperature. The Dye-loaded neutrophils were stimulated with PMA (2ng/mL). Fluorescence images were acquired before and 10 minutes after stimulation. Excitation: 488nm emission: 505-550 nm barrier filters
ImmunoChemistry Technologies Highly Reactive Oxygen Species (hROS) Detection Kit
$304.55
Figure 1. Detection of Hypochlorite ( -OCl ) in neutrophils. Neutrophils were isolated from porcine blood, washed in Krebs-Ringers phosphate buffer (as described in step V 1. above) and seeded in glass bottom dishes. The cells were then loaded with APF or HPF (10 mM final) by incubation for 30 minutes at room temperature. The Dye-loaded neutrophils were stimulated with PMA (2ng/mL).Figure 2. Fluorescence images were acquired before and 10 minutes after stimulation. Excitation: 488 nm emission: 505-550 nm barrier filters. Hypochlorite production can be detected with increase APF fluorescence and no increase in HPF fluorescence.
ImmunoChemistry Technologies Fluorescent Hypochlorite Detection Kit
$533.50
Figure 1. Jurkat cells were stimulated with staurosporine (B) or DMSO (A), then stained according to the kit protocol. The cells were washed twice and analyzed by flow cytometry: Ex:488nm Em: FL1 and FL2. A. Healthy cells show a strong red fluorescence (intact mitochondria) and no green fluorescence (no active caspases). B. Apoptotic cells show a loss of red fluorescence (y axis) due to loss of mitochondrial membrane potential and positive green fluorescence (x axis) indicating active caspases.Figure 2. HeLa Cells activated with staurosporine (B) or DMSO (A), then stained according to the kit protocol. The cells were washed twice and analyzed by flow cytometry: Ex:488nm Em: FL1 and FL2. A. Healthy cells show a strong red fluorescence (intact mitochondria) and no green fluorescence (no active caspases). B. Cells becoming apoptotic show loss of red fluorescence (y axis) due to loss of mitochondrial membrane potential and positive green fluorescence (x axis) indicating active caspases.
ImmunoChemistry Technologies Dual Caspase 1 and Mitochondrial Membrane Potential Assay Kit
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Figure 1. Jurkat cells were stimulated with staurosporine (B) or DMSO (A), then stained according to the kit protocol. The cells were washed twice and analyzed by flow cytometry: Ex:488nm Em: FL1 and FL2. A. Healthy cells show a strong red fluorescence (intact mitochondria) and no green fluorescence (no active caspases). B. Apoptotic cells show a loss of red fluorescence (y axis) due to loss of mitochondrial membrane potential and positive green fluorescence (x axis) indicating active caspases.Figure 2. HeLa Cells activated with staurosporine (B) or DMSO (A), then stained according to the kit protocol. The cells were washed twice and analyzed by flow cytometry: Ex:488nm Em: FL1 and FL2. A. Healthy cells show a strong red fluorescence (intact mitochondria) and no green fluorescence (no active caspases). B. Cells becoming apoptotic show loss of red fluorescence (y axis) due to loss of mitochondrial membrane potential and positive green fluorescence (x axis) indicating active caspases.
ImmunoChemistry Technologies Dual Caspase 3/7 and Mitochondrial Membrane Potential Assay Kit
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Figure 1. Jurkat cells were stimulated with staurosporine (B) or DMSO (A), then stained according to the kit protocol. The cells were washed twice and analyzed by flow cytometry: Ex:488nm Em: FL1 and FL2. A. Healthy cells show a strong red fluorescence (intact mitochondria) and no green fluorescence (no active caspases). B. Apoptotic cells show a loss of red fluorescence (y axis) due to loss of mitochondrial membrane potential and positive green fluorescence (x axis) indicating active caspases.Figure 2. HeLa Cells activated with staurosporine (B) or DMSO (A), then stained according to the kit protocol. The cells were washed twice and analyzed by flow cytometry: Ex:488nm Em: FL1 and FL2. A. Healthy cells show a strong red fluorescence (intact mitochondria) and no green fluorescence (no active caspases). B. Cells becoming apoptotic show loss of red fluorescence (y axis) due to loss of mitochondrial membrane potential and positive green fluorescence (x axis) indicating active caspases.
ImmunoChemistry Technologies Dual Poly Caspase and Mitochondrial Membrane Potential Assay Kit
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Figure 1. Jurkat Cells were stimulated with DMSO for 2 hours. The cells were washed and fixed for 15 minutes. The cells were then permeabilized with saponin and stained with Rabbit anti-active caspase 3 antibody for 1 hour. The samples were then stained with PE labeled Goat anti-Rabbit antibody.Figure 2. Jurkat Cells were stimulated with staurosporine for 2 hours. The cells were washed and fixed for 15 minutes. The cells were then permeabilized with saponin and stained with Rabbit anti-active caspase 3 antibody for 1 hour. The samples were then stained with PE labeled Goat anti-Rabbit antibody.
ImmunoChemistry Technologies Apo Active PE - Caspase 3 Detection Kit
$533.50
Figure 1. U937 cells were stimulated with staurosporine for 5 hours, and then assayed for caspase 3/7 activity following the kit protocol.
ImmunoChemistry Technologies Apo 3/7 HTS Assay Kit
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Figure 1. Assay principle showing the effect of inhibitorsFigure 2. Assay principle schematic
ImmunoChemistry Technologies Bioluminescent Acetylcholinesterase Assay Kit
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Figure 1. Balb/c mice were either inoculated with HSV-1 virus, or given a sham treatment. Seven days later, the mice were injected IV with either DyLight® 747-VAD-FMK (kit 9114), NIR-DyLight® 747 Free Dye (kit 9115), or no reagent. Strong caspase activity was seen in the brain of the HSV-1 infected / NIR-DyLight® 747 Tracer-treated mouse. The liver is the route of clearance for DyLight® Tracers. All mice show fluorescence signal in the tail where reagent is likely to pool after IV injection.
ImmunoChemistry Technologies Near Infrared DyLight® 747 Free Dye Control Assay
$318.90
Figure 1. Apoptotic tissues in murine tumor models were imaged in vivo with 747-VAD-FMK (Cat. 9114). Negative control (placebo) mice (left) and experimentally treated mice (right) were injected intravenously with DyLight® 747-VAD-FMK tracer and imaged non-invasively at various time points with the CRi Maestro™ imaging system. Apoptotic tumor tissues fluoresce bright blue in the experimental group subjects, indicating that the experimental treatment had an apoptotic effect.
ImmunoChemistry Technologies Near-Infrared DyLight® 747 Tracer in vivo Active Caspase (VAD) Assay
$461.95
Figure 1. Brain abscesses were induced in mice following inoculation of live S. aureus. Mice received i.v. injections of DyLight® 690-VAD-FMK tracer or DyLight® 690 Dye Control (Cat. 9113) 17 hours post-infection; brain tissue was imaged 1 hour later ex vivo using an IVIS® Spectrum™ (Caliper Life Sciences). Strong caspase activity was seen in brain abscesses using DyLight® 690-VAD-FMK tracer (right); minimal signal was detected in the DyLight® 690 Free Dye Control (left).
ImmunoChemistry Technologies Near Infrared DyLight® 690 Free Dye Control Assay
$318.90
Figure 1. Brain abscesses were induced in mice following inoculation of live S. aureus. Mice received i.v. injections of DyLight® 690-VAD-FMK tracer or DyLight® 690 Dye Control (Cat. 9113) 17 hours post-infection; brain tissue was imaged 1 hour later ex vivo using an IVIS® Spectrum™ (Caliper Life Sciences). Strong caspase activity was seen in brain abscesses using DyLight® 690-VAD-FMK tracer (right); minimal signal was detected in the DyLight® 690 Dye Control Free Dye control (left).Experimental design of Aim2WT or Aim2−/− bone marrow (BM) transplantation to Ldlr−/−:Mx1cre:c-mybfl/fl mice receiving i.v. DNA (top) and flow cytometry histograms (bottom) of caspase-1 activity(Assay kit, Cat no. 9122) in bone marrow recipients 24 h after i.v. DNA. Image from publication, CC-BY-4.0, PMID: 39112714.
ImmunoChemistry Technologies Near-Infrared DyLight® 690 Tracer in vivo Active Caspase (VAD) Assay
$461.95
Figure 1. Excitation and emission spectra. Red Fluorescent SR In vivo Poly (active) Caspase (VAD) Assay inhibitor probe was reconstituted in DMSO, diluted in an aqueous buffer, and then analyzed on a Molecular Devices M5e plate reader. The excitation spectrum (black) was generated using an emission of 630 nm. The emission spectrum (orange) was generated using an excitation of 540 nm. The probe optimally excites at 550-580 nm and has a peak emission at 590-600 nm.Figure 2. Mice were given either a 0Gy or 15Gy whole lung dose of gamma irradiation. SR in vivo probe (Cat. 982/983) was injected and allowed to circulate for 18 hours prior to sacrifice. Paraffin sections were prepared, nuclei stained with DAPI (blue), and lungs imaged with a fluorescence microscope. Caspase activity (red) was induced in irradiated mice compared to control animals. Data courtesy of E. Hernady (University of Rochester Medical Center, Rochester, NY).
ImmunoChemistry Technologies Red Fluorescent SR In vivo Poly (active) Caspase (VAD) Assay
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Figure 1. Green Fluorescent FAM In vivo Poly (active) Caspase (VAD) Assay inhibitor probe was reconstituted in DMSO, diluted in an aqueous buffer, and then analyzed on a Molecular Devices M5e plate reader. The excitation spectrum (black) was generated using an emission of 590 nm. The emission spectrum (green) was generated using an excitation of 460 nm.Figure 2. Staurosporine (Cat. 6212) was injected into the forebrain of a female house sparrow to induce apoptosis. Approximately 20 hours later, FAM in vivo probe (Cat. 980/981) was intravenously injected into the jugular and allowed to circulate 30 minutes. The bird was sacrificed and frozen brain sections prepared. Neurons with active caspases fluoresce green. This image shows one apoptotic neuron at 100X. Data courtesy of Mr. Chris Thompson at the University of Washington, Seattle.
ImmunoChemistry Technologies Green Fluorescent FAM In vivo Poly (active) Caspase (VAD) Assay
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Figure 1. Jurkat cells were treated with a negative control (left) or staurosporine (right), then stained using Swine Annexin V-Fluorescein Assay (Cat. 9142). Annexin V-Fluorescein labeled only 13.1% of control cells (UR + LR = 11.0% + 3.1%, left), whereas staurosporine induced Annexin V-Fluorescein labeling in 96.7% of cells (UR + LR = 32.7% + 64.0%, right). Staurosporine also significantly increased the proportion of dually stained cells (UR). Data courtesy of Mrs. T. Murphy, ICT, 227:19.
ImmunoChemistry Technologies Swine Annexin V-Fluorescein Apoptosis Assay Kit
$479.35
Figure 1. Jurkat cells were treated with a negative control (left) or staurosporine (right), washed, then stained with Equine Annexin V-Fluorescein Assay (Cat. 9141). Equine Annexin V-Fluorescene labeled only 17.4% of the control cells (UR + LR = 10.3% + 7.1%, left); staurosporine induced Equine Annexin V-Fluorescene labeling in 97.6% of cells (UR + LR = 27.8% + 69.8%, right). Staurosporine increased the proportion of dual stained cells (UR). Data courtesy of Mrs. T. Murphy, ICT, 227:68.
ImmunoChemistry Technologies Equine Annexin V-Fluorescein Apoptosis Assay Kit
$479.35
Figure 1. Jurkat cells were treated with a negative control (left) or staurosporine (right), washed, then stained with Chicken Annexin V-Fluorescein Assay (Cat. 9140). Chicken Annexin V-Fluorescein labeled only 16.9% of control cells (UR + LR = 13.2% + 3.7%, left); staurosporine induced Chicken Annexin V-Fluorescein labeling in 97% of cells (UR + LR = 46.6% + 50.4%, right). Staurosporine increased the proportion of dual stained cells (UR). Data courtesy of Mrs. Tracy Murphy, ICT, 227:19.
ImmunoChemistry Technologies Chicken Annexin V-Fluorescein Apoptosis Assay Kit
$479.35
Figure 1. Jurkat cells were treated with a negative control (left) or staurosporine (right), then stained with Canine Annexin V-Fluorescein Assay (Cat. 9139). Annexin V-Fluorescein labeled only 15.6% of the control cells (UR + LR = 9.8% + 5.8%, left); staurosporine induced Annexin V-Fluorescein labeling in 97.3% of cells (UR + LR = 11.5% + 85.8%, right). Staurosporine did not significantly increase the proportion of dually stained cells (UR). Data courtesy of Mrs. T. Murphy, ICT, 224:69-71.
ImmunoChemistry Technologies Canine Annexin V-Fluorescein Apoptosis Assay Kit
$479.35
Figure 1. Jurkat cells were treated with a negative control (left) or staurosporine (right), washed, and then stained with Bovine Annexin V-Fluorescein (Cat. 9138). Bovine Annexin V-Fluorescein labeled only 17.6% of control cells (UR + LR = 13.5% + 4.1%, left); staurosporine induced Bovine Annexin V-Fluorescein labeling in 98.8% of cells (UR + LR = 15.3% + 83.5%, right). Staurosporine did not increase the proportion of dual stained cells (UR). Data courtesy of Mrs. Tracy Hanson, ICT, 224:69-71.
ImmunoChemistry Technologies Bovine Annexin V-Fluorescein Apoptosis Assay Kit
$479.35
Figure 1. Quantification of 4 populations in each sampleFigure 2. Assay procedure
ImmunoChemistry Technologies Cell-mediated Cytotoxicity Assay: Total Cytotoxicity Test
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Native Human Cathepsin D
ImmunoChemistry Technologies Native Human Cathepsin D
$364.30
ImmunoChemistry Technologies Cathepsin B
$355.75

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