Caspases

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Figure 1. THP-1 cells were treated with either a negative control (A), or PMA (5 ng/mL) followed by LPS (10 ng/mL). Cells were then stained with FAM-YVAD-FMK (cat. 97/98) and analyzed by phase contrast and fluorescence microscopy. In the treated sample, many cells appear bright green, indicating increased caspase-1 activity (B). In the non-induced sample, few green cells are visible, indicating a low level of caspase-1 activity (A). Data courtesy of Dr. Brian Lee, ICT (207:11).
ImmunoChemistry Technologies Green Fluorescent FAM-FLICA® Caspase-1 (YVAD) Assay Kit
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Figure 1. Signals leading to activation of the NLRP3 inflammasome.Figure 2. C2BBe1 cells were infected with wild type Salmonella constitutively expressing mCherry (red). After 9 hours, live cells were incubated with FAM-YVAD-FMK (green cell) for 1 hour. The infected cell (red) is undergoing pyroptosis, and shows increased green fluorescence compared to uninfected cells due to caspase-1 activation. Data courtesy Knodler, Leigh A., et al. Dissemination of invasive Salmonella via bacterial-induced extrusion of mucosal epithelia. PNAS. 107:41, 17733-17738 (2010).
ImmunoChemistry Technologies Pyroptosis/Caspase-1 Assay - Green Fluorescent
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Figure 1. FLICA® 660 was reconstituted in DMSO, diluted in diH2O, and analyzed in a fluorescence plate reader. The excitation spectrum was generated using an emission of 760 nm. The emission spectrum was generated using an excitation of 600 nm.Figure 2. Flow cytometry analysis of Jurkat cells treated with FLICA® 660-YVAD-FMK. After spiking FLICA® into the cell culture media, the cells were treated with either a negative control (left) or nigericin (20 μM, no.6698) to induce caspase-1 activity (right) for 24 hours. The negative control induced caspase-1 activity in 6.8% of cells (left); treatment with nigericin induced caspase-1 activity in 57.3% of cells (right). This is a ratio of 8:1. Data courtesy of Mrs. Tracy Murphy, ICT, 230-01.
ImmunoChemistry Technologies Far-Red Fluorescent FLICA® 660 Caspase-1 (YVAD) Assay Kit
$265
Figure 1. Flow cytometry analysis of active caspase-9 in MDA-MB-231 (A,B) and ZR-75-1 (C,D) cells. Cells were stained with FAM-FLICA® Caspase-9 reagent. From Figure 5 of R. Krętowski and M. Cechowska-Pasko, The Reduced Graphene Oxide (rGO) Induces Apoptosis, Autophagy and Cell Cycle Arrest in Breast Cancer Cells, Int J Mol Sci (2022) 23, 9285. Used under the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). Copyright © 2022 by the authors.Flow cytometry analysis of the caspase-9 activity (Assay Kit, Cat no. 912) in the MCF-7 (A,B) and MDA-MB-231 (C,D) breast cancer cells after 24 h of incubation with Les-6287 (1.0 μM and 1.5 μM), doxorubicin (1.0 μM), and DMSO. Image from publication, CC-BY-4.0, PMID: 39199694.
ImmunoChemistry Technologies Green Fluorescent FAM-FLICA® Caspase-9 (LEHD) Assay Kit
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Figure 1. Monocytes can secrete active caspase-1, therefore cells were labeled with FAM-FLICA® Caspase-1 (WEHD) (cat. 9161/9162) prior to treatment. Cells were then treated with DMSO (control) or 10 µM nigericin (cat. 6698), stained with Hoechst 33342, fixed (cat. 636), and viewed using EGFP (Ex 470/30, Em 530/50) and DAPI (Ex 375/28, Em 460/50) LED filter cubes at 20X. Increased levels of FAM-FLICA® staining were seen in the nigericin-treated cells. Data courtesy of Dr. Kristi Strandberg, ICT (235:60).
ImmunoChemistry Technologies Green Fluorescent FAM FLICA® Caspase-1 (WEHD) Assay Kit
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Figure 1. Suspension Jurtkat cells were treated with DMSO (control, A) or 1 µM staurosporine (B) for 4 hours at 37°C, then stained with SR-LETD-FMK (kit cat. 9149/9150) for 1 hour at 37°C. Cells were washed three times and slides prepared. Treated cells appear bright red, indicating a high level of caspase-8 activity (B). Few non-induced cells are red, indicating minimal caspase-8 activity (A, Non-Induced, left). Data courtesy of Mrs. Tracy Murphy, ICT (220:68, 121815).
ImmunoChemistry Technologies Red Fluorescent SR-FLICA® Caspase-8 (LETD) Assay Kit
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Figure 1. Cresyl Violet Perchlorate Excitation and Emission Spectra in EthanolFigure 2. MCF-7 cells were exposed to 0.15 µM camptothecin (cat. 6210) for 24 hours at 37°C, then stained with MR-(DEVD)2 for 30 minutes at 37°C, washed, and then stained with 1 µg/mL Hoechst stain. Apoptotic cells with orange-red lysosomal bodies and less intense blue nuclei are intermixed with non-apoptotic cells bearing bright blue nuclei and absent or reduced orange-red lysosomal staining. Photo provided by Dr. Zbigniew Darzynkiewicz at Brander Cancer Research Center Institute, New York, NY.
ImmunoChemistry Technologies Magic Red® Fluorescent Caspase-3/7 Assay Kit
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Figure 1. Cell death in primary rat hippocampal neurons. (A) is a composite of FLICA® FAM-DEVD-FMK (B), PI (C), and Hoechst (D). DNA is labeled in 4 cells (D), 3 cells are caspase-positive (B), 2 are membrane-compromised (C). One cell is in early apoptosis (green but not red). Two FLICA®-positive cells are also PI-positive (red, C) (becoming membrane compromised) and are in the late stages of apoptosis rather than necrosis. Data courtesy of Dr. Z. Kahraman Akozer, University of Maryland.Figure 2. Normal (A) and keratoconus (B) corneal fibroblasts were treated with 200 μM H2O2 for 1 hour, washed, and allowed to recover. Cells were then treated with FAM-DEVD-FMK (cat. 93/94) for 1 hour, then washed with 1X Apoptosis Wash Buffer. Keratoconus corneal fibroblasts treated with H2O2 (B) show a significant increase in caspase-3/7 activity compared to normal cells (A). Non-apoptotic cells are dark in background. Data courtesy of Dr. Cristina Kenney, University of California, Irvine.
ImmunoChemistry Technologies Green Fluorescent FAM-FLICA® Caspase-3/7 (DEVD) Assay Kit
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Figure 1. FLICA® 660 was reconstituted in DMSO, diluted in diH2O, and analyzed in a fluorescence plate reader. The excitation spectrum was generated using an emission of 760 nm. The emission spectrum was generated using an excitation of 600 nm.Figure 2. Jurkat cells were treated with a negative control (left) or staurosporine, an apoptosis-inducing agent (right), for 4 hours, then stained with FLICA® 660-DEVD-FMK (cat. 9125) for 1 hour. Cells were washed and analyzed by flow cytometer. The negative control induced caspase-3/7 activity in 4.4% of cells (left); treatment with staurosporine induced caspase-3/7 activity in 70.9% of cells (right). This is a ratio of 16:1. Data courtesy of Mrs. Tracy Murphy, ICT, 13F38, 022013.
ImmunoChemistry Technologies Far-Red Fluorescent FLICA® 660 Caspase-3/7 (DEVD) Assay Kit
$264
Figure 1. Control Alzheimer's Disease (AD) macrophages and curcumin-treated AD macrophages were exposed to FITC-labeled Aß and stained with SR-DEVD-FMK (#931/932). Macrophages that engulf FITC-Aß fluoresce green (right). Caspase-positive cells fluoresce red (left). Essentially all of the untreated AD macrophages engulfed FITC-Aß and became apoptotic. Curcumin-treated AD macrophages engulfed FITC-Aß but did not become apoptotic. Data courtesy of Dr. Milan Fiala, UCLA 072205.
ImmunoChemistry Technologies Red Fluorescent SR-FLICA® Caspase-3/7 (DEVD) 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 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
$522
Figure 1. Jurkat Cells were stimulated with staurosporine (B) or DMSO (A) for 2 hours. The cells were washed and fixed for 15 minutes. The cells were then permeabilized with saponin and stained with anti-active caspase 3 antibody for 1 hour. The samples were then stained with FITC labeled Goat anti-Rabbit antibody.
ImmunoChemistry Technologies Apo Active FITC - Caspase 3 Detection Kit
$522
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. 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
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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
$452
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
$312
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
$452
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 staurosporine (Positive, Induced, right histogram). Negative control cells were spiked with a DMSO vehicle control (Negative, Non-induced, left histogram). Cells were stained with SR-VAD-OPH for 1 hour, washed twice, and read on a flow cytometer. Only 8.0% of control cells displayed caspase activity, whereas staurosporine induced caspase activity in 90.3% of the experimental cells. This is a ratio of 11:1. Data courtesy of Ms. Tracy Hanson, ICT 092713.Figure 2. Jurkat cells were treated with staurosporine to induce apoptosis, then labeled with SR-VAD-OPH for 60 minutes. Cells were washed twice and wet-mount slides were prepared. A grey DIC image (left) was taken, and red fluorescence (right) was detected using a band pass filter (excitation at 565 nm, emission at 590-600 nm). SR-VAD-OPH reveals complete caspase activation in the form of red fluorescence in all cells. Data courtesy of Ms. Tracy Hanson, ICT (ICT111809; SR_OPH_111_17_18).
ImmunoChemistry Technologies Red Fluorescent SR-VAD-OPH in vitro Poly Caspase Apoptosis Detection Reagent
$572
Figure 1. Jurkat cells were treated with staurosporine to induce apoptosis, and then labeled with FAM-DEVD-OPH (cat. 6356). Wet-mount slides were prepared and green fluorescence (right) was detected using a band pass filter (excitation at 488 nm, emission at 520 nm). FAM-DEVD-OPH reveals complete caspase-3/7 activation in the form of green fluorescence in all cells. The corresponding DIC image is shown (left). Data courtesy of Ms. Tracy Hanson, ICT (ICT111709; FAM-DEVD-OPH_in_59_60).
ImmunoChemistry Technologies Green Fluorescent FAM-DEVD-OPH in vitro caspase-3/7 Apoptosis Detection Reagent
$572

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