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Cell Death Detection ELISA PLUS roche详细信息：
Application
The Cell Death Detection ELISAPLUS photometric enzyme immunoassay is used for the quantitative in vitro determination of cytoplasmic histone-associated DNA fragments (mono- and oligonucleosomes) after induced cell death.
The kit contains a stop solution which allows users to terminate the substrate reaction and run the assay under defined conditions, making it suitable for use in high-throughput applications.
Benefits
One-step ELISA
High sensitivity: (5 x 102 cells/ml)
Fast performance: (3 - 4 hours)
Positive control included
No prelabeling of cells necessary
Nonradioactive assay system
No species restriction
Easy handling
Low background
Suppressed human anti-mouse factor
Function-tested
Product Description
Cell Death Detection ELISA PLUS rocheAssay time:
3 - 4 hours
Negative control:
Depending on cell culture conditions, each exponentially growing permanent cell culture contains a certain amount of dead cells (typically approximately 3 - 8%). In the immunoassay, these inherent dead cells in the untreated sample (without a cell-death-inducing agent) will cause a certain absorbance value (negative control).
Positive control:
A DNA-histone complex serves as a positive control.
Sample material:
Cytoplasmic fractions (lysates) of cell lines, cells ex vivo, cell culture supernatants, and serum or plasma
Sensitivity:
The exact detection limit of dying/dead cells in a particular sample strongly depends on the kinetics of cell death, the cytotoxic agent used, and the amount of affected cells in the total cell population. Using U937/camptothecin (CAM) as a cellular model system for cell death, the immunoassay allows the specific detection of mono- and oligonucleosomes in the cytoplasmic fraction of 125 cell equivalents/well.
Specificity:
Anti-histone reacts with the histones H1, H2A, H2B, H3, and H4 of various species (e.g., human, mouse, rat, hamster, cow, opossum, Xenopus). Anti-DNA binds to single- and double-stranded DNA. Therefore, the ELISA allows the detection of mono- and oligonucleosomes from various species, and may be applied to measure apoptotic cell death in many different cell systems.
Cell Death Detection ELISA PLUS rocheBackground Information
Two distinct forms of eukaryotic cell death can be classified by morphological and biochemical criteria: necrosis and apoptosis. Necrosis is accompanied by increased ion permeability of the plasma membrane; the cells swell and the plasma membrane ruptures within minutes (osmotic lysis). Apoptosis is characterized by membrane blebbing (zeiosis), condensation of cytoplasm, and the activation of an endogenous endonuclease. This Ca2+- and Mg2+-dependent nuclease cleaves double-stranded DNA at the most accessible internucleosomal linker region, generating mono- and oligonucleosomes. In contrast, the DNA of the nucleosomes is tightly complexed with the core histones H2A, H2B, H3, and H4, and is thus protected from cleavage by the endonuclease. The yielded DNA fragments are discrete multiples of an 180-bp subunit, detected as a “DNA ladder” on agarose gels after extraction and separation of the fragmented DNA. The enrichment of mono- and oligonucleosomes in the cytoplasm of the apoptotic cell is due to the fact that DNA degradation occurs several hours before plasma membrane breakdown.
Contents
Anti-histone-biotin (clone H11-4)
Anti-DNA-POD (clone MCA-33)
Positive Control
Incubation Buffer
Lysis Buffer
Substrate Buffer
ABTS Substrate Tablet
ABTS Stop Solution
Microplate (streptavidin-coated)
Adhesive Cover Foils
Principle
The assay is based on the quantitative “sandwich enzyme immunoassay” principle using mouse monoclonal antibodies directed against DNA and histones. This allows the specific determination of mono- and oligonucleosomes in the cytoplasmic fraction of cell lysates. The samples are placed into a streptavidin-coated microplate and incubated with a mixture of anti-histone-biotin and anti-DNA-peroxidase. During the incubation interval, nucleosomes will be captured via their histone component by the anti-histone-biotin antibody, while binding to the streptavidin-coated microplate. Simultaneously, anti-DNA-peroxidase binds to the DNA part of the nucleosomes. After removal of the unbound antibodies, the amount of peroxidase retained in the immunocomplex is photometrically determined with ABTS as the substrate.
Cell Death Detection ELISA PLUS roche
xy1201 HOMEC 人卵巢微血管内皮细胞 Human Ovarian Microvascular Endothelial Cells 5 x 10^5 cells/vial ATCC
xy1202 HOSEpiC 人卵巢表层上皮细胞 Human Ovarian Surface Epithelial Cells 5 x 10^5 cells/vial ATCC
xy1203 HOF 人卵巢成纤维细胞 Human Ovarian Fibroblasts 5 x 10^5 cells/vial ATCC
xy1204 HMMEC 人乳腺微血管内皮细胞 Human Mammary Microvascular Endothelial Cells 5 x 10^5 cells/vial ATCC
xy1205 HMEpiC 人乳腺上皮细胞 Human Mammary Epithelial Cells 5 x 10^5 cells/vial ATCC
xy1206 HMF 人乳腺成纤维细胞 Human Mammary Fibroblasts 5 x 10^5 cells/vial ATCC
xy1207 ATCC
xy1208 间充质干细胞系统 ATCC
xy1209 HAMSC 人羊膜间充质基质细胞 Human Amniotic Mesenchymal Stromal Cells 5 x 10^5 cells/vial ATCC
xy1210 HCMSC 人绒毛膜间充质基质细胞 Human Chorionic Mesenchymal Stromal Cells 5 x 10^5 cells/vial ATCC
xy1211 HMSC-bm 人间充质干细胞--骨髓 Human Mesenchymal Stem Cells-bone marrow 5 x 10^5 cells/vial ATCC
xy1212 HMSC-ad 人间充质干细胞--脂肪 Human Mesenchymal Stem Cells-adipose 5 x 10^5 cells/vial ATCC
xy1213 HMSC-hp 人间充质干细胞--肝脏 Human Mesenchymal Stem Cells-hepatic 5 x 10^5 cells/vial ATCC
xy1214 HUMSC 人脐带间充质干细胞 Human Umbilical Mesenchymal Stem Cells 5 x 10^5 cells/vial ATCC
xy1215 HPMSC 人肺间充质干细胞 Human Pulmonary Mesenchymal Stem Cells 5 x 10^5 cells/vial ATCC
xy1216 HVMSC 人脊椎间充质干细胞 Human Vertebral Mesenchymal Stem Cells 5 x 10^5 cells/vial ATCC
xy1217 ATCC
xy1218 ATCC
xy1219 脐带细胞系统 ATCC
xy1220 HUVEC 人脐静脉内皮细胞 Human Umbilical Vein Endothelial Cells 5 x 10^5 cells/vial ATCC
xy1221 HUAEC 人脐动脉内皮细胞 Human Umbilical Artery Endothelial Cells 5 x 10^5 cells/vial ATCC
xy1222 HUVSMC 人脐静脉平滑肌细胞 Human Umbilical Vein Smooth Muscle Cells 5 x 10^5 cells/vial ATCC
xy1223 HUASMC 人脐动脉平滑肌细胞 Human Umbilical Artery Smooth Muscle Cells 5 x 10^5 cells/vial ATCC
xy1224 ATCC
xy1225 小鼠细胞系统 ATCC
xy1226 MPC 小鼠垂体细胞 Mouse Pituitary Cells 5 x 10^5 cells/vial ATCC
xy1227 MMenC 小鼠脑膜细胞 Mouse Meningeal Cells 5 x 10^5 cells/vial ATCC
xy1228 MN-r 小鼠脑脊神经元 Mouse Neurons-raphe 1 x 10^6 cells/vial ATCC
xy1229 MN-c 小鼠皮质神经元 Mouse Neurons-cortical 1 x 10^6 cells/vial ATCC
xy1230 MGC 小鼠颗粒细胞 Mouse Granule Cells 1 x 10^6 cells/vial ATCC
xy1231 MN-h 小鼠海马趾神经元 Mouse Neurons-hippocampal 1 x 10^6 cells/vial ATCC
xy1232 MN-sn 小鼠黑质神经元 Mouse Neurons-substantia nigra 1 x 10^6 cells/vial ATCC
xy1233 MN-s 小鼠纹状体神经元 Mouse Neurons-striatal 1 x 10^6 cells/vial ATCC
xy1234 MN-vsc 小鼠腹脊髓神经元 Mouse Neurons-ventral spinal cord 1 x 10^6 cells/vial ATCC
Application
The Cell Death Detection ELISAPLUS photometric enzyme immunoassay is used for the quantitative in vitro determination of cytoplasmic histone-associated DNA fragments (mono- and oligonucleosomes) after induced cell death.
The kit contains a stop solution which allows users to terminate the substrate reaction and run the assay under defined conditions, making it suitable for use in high-throughput applications.
Benefits
One-step ELISA
High sensitivity: (5 x 102 cells/ml)
Fast performance: (3 - 4 hours)
Positive control included
No prelabeling of cells necessary
Nonradioactive assay system
No species restriction
Easy handling
Low background
Suppressed human anti-mouse factor
Function-tested
Product Description
Cell Death Detection ELISA PLUS rocheAssay time:
3 - 4 hours
Negative control:
Depending on cell culture conditions, each exponentially growing permanent cell culture contains a certain amount of dead cells (typically approximately 3 - 8%). In the immunoassay, these inherent dead cells in the untreated sample (without a cell-death-inducing agent) will cause a certain absorbance value (negative control).
Positive control:
A DNA-histone complex serves as a positive control.
Sample material:
Cytoplasmic fractions (lysates) of cell lines, cells ex vivo, cell culture supernatants, and serum or plasma
Sensitivity:
The exact detection limit of dying/dead cells in a particular sample strongly depends on the kinetics of cell death, the cytotoxic agent used, and the amount of affected cells in the total cell population. Using U937/camptothecin (CAM) as a cellular model system for cell death, the immunoassay allows the specific detection of mono- and oligonucleosomes in the cytoplasmic fraction of 125 cell equivalents/well.
Specificity:
Anti-histone reacts with the histones H1, H2A, H2B, H3, and H4 of various species (e.g., human, mouse, rat, hamster, cow, opossum, Xenopus). Anti-DNA binds to single- and double-stranded DNA. Therefore, the ELISA allows the detection of mono- and oligonucleosomes from various species, and may be applied to measure apoptotic cell death in many different cell systems.
Cell Death Detection ELISA PLUS rocheBackground Information
Two distinct forms of eukaryotic cell death can be classified by morphological and biochemical criteria: necrosis and apoptosis. Necrosis is accompanied by increased ion permeability of the plasma membrane; the cells swell and the plasma membrane ruptures within minutes (osmotic lysis). Apoptosis is characterized by membrane blebbing (zeiosis), condensation of cytoplasm, and the activation of an endogenous endonuclease. This Ca2+- and Mg2+-dependent nuclease cleaves double-stranded DNA at the most accessible internucleosomal linker region, generating mono- and oligonucleosomes. In contrast, the DNA of the nucleosomes is tightly complexed with the core histones H2A, H2B, H3, and H4, and is thus protected from cleavage by the endonuclease. The yielded DNA fragments are discrete multiples of an 180-bp subunit, detected as a “DNA ladder” on agarose gels after extraction and separation of the fragmented DNA. The enrichment of mono- and oligonucleosomes in the cytoplasm of the apoptotic cell is due to the fact that DNA degradation occurs several hours before plasma membrane breakdown.
Contents
Anti-histone-biotin (clone H11-4)
Anti-DNA-POD (clone MCA-33)
Positive Control
Incubation Buffer
Lysis Buffer
Substrate Buffer
ABTS Substrate Tablet
ABTS Stop Solution
Microplate (streptavidin-coated)
Adhesive Cover Foils
Principle
The assay is based on the quantitative “sandwich enzyme immunoassay” principle using mouse monoclonal antibodies directed against DNA and histones. This allows the specific determination of mono- and oligonucleosomes in the cytoplasmic fraction of cell lysates. The samples are placed into a streptavidin-coated microplate and incubated with a mixture of anti-histone-biotin and anti-DNA-peroxidase. During the incubation interval, nucleosomes will be captured via their histone component by the anti-histone-biotin antibody, while binding to the streptavidin-coated microplate. Simultaneously, anti-DNA-peroxidase binds to the DNA part of the nucleosomes. After removal of the unbound antibodies, the amount of peroxidase retained in the immunocomplex is photometrically determined with ABTS as the substrate.
Cell Death Detection ELISA PLUS roche
xy1201 HOMEC 人卵巢微血管内皮细胞 Human Ovarian Microvascular Endothelial Cells 5 x 10^5 cells/vial ATCC
xy1202 HOSEpiC 人卵巢表层上皮细胞 Human Ovarian Surface Epithelial Cells 5 x 10^5 cells/vial ATCC
xy1203 HOF 人卵巢成纤维细胞 Human Ovarian Fibroblasts 5 x 10^5 cells/vial ATCC
xy1204 HMMEC 人乳腺微血管内皮细胞 Human Mammary Microvascular Endothelial Cells 5 x 10^5 cells/vial ATCC
xy1205 HMEpiC 人乳腺上皮细胞 Human Mammary Epithelial Cells 5 x 10^5 cells/vial ATCC
xy1206 HMF 人乳腺成纤维细胞 Human Mammary Fibroblasts 5 x 10^5 cells/vial ATCC
xy1207 ATCC
xy1208 间充质干细胞系统 ATCC
xy1209 HAMSC 人羊膜间充质基质细胞 Human Amniotic Mesenchymal Stromal Cells 5 x 10^5 cells/vial ATCC
xy1210 HCMSC 人绒毛膜间充质基质细胞 Human Chorionic Mesenchymal Stromal Cells 5 x 10^5 cells/vial ATCC
xy1211 HMSC-bm 人间充质干细胞--骨髓 Human Mesenchymal Stem Cells-bone marrow 5 x 10^5 cells/vial ATCC
xy1212 HMSC-ad 人间充质干细胞--脂肪 Human Mesenchymal Stem Cells-adipose 5 x 10^5 cells/vial ATCC
xy1213 HMSC-hp 人间充质干细胞--肝脏 Human Mesenchymal Stem Cells-hepatic 5 x 10^5 cells/vial ATCC
xy1214 HUMSC 人脐带间充质干细胞 Human Umbilical Mesenchymal Stem Cells 5 x 10^5 cells/vial ATCC
xy1215 HPMSC 人肺间充质干细胞 Human Pulmonary Mesenchymal Stem Cells 5 x 10^5 cells/vial ATCC
xy1216 HVMSC 人脊椎间充质干细胞 Human Vertebral Mesenchymal Stem Cells 5 x 10^5 cells/vial ATCC
xy1217 ATCC
xy1218 ATCC
xy1219 脐带细胞系统 ATCC
xy1220 HUVEC 人脐静脉内皮细胞 Human Umbilical Vein Endothelial Cells 5 x 10^5 cells/vial ATCC
xy1221 HUAEC 人脐动脉内皮细胞 Human Umbilical Artery Endothelial Cells 5 x 10^5 cells/vial ATCC
xy1222 HUVSMC 人脐静脉平滑肌细胞 Human Umbilical Vein Smooth Muscle Cells 5 x 10^5 cells/vial ATCC
xy1223 HUASMC 人脐动脉平滑肌细胞 Human Umbilical Artery Smooth Muscle Cells 5 x 10^5 cells/vial ATCC
xy1224 ATCC
xy1225 小鼠细胞系统 ATCC
xy1226 MPC 小鼠垂体细胞 Mouse Pituitary Cells 5 x 10^5 cells/vial ATCC
xy1227 MMenC 小鼠脑膜细胞 Mouse Meningeal Cells 5 x 10^5 cells/vial ATCC
xy1228 MN-r 小鼠脑脊神经元 Mouse Neurons-raphe 1 x 10^6 cells/vial ATCC
xy1229 MN-c 小鼠皮质神经元 Mouse Neurons-cortical 1 x 10^6 cells/vial ATCC
xy1230 MGC 小鼠颗粒细胞 Mouse Granule Cells 1 x 10^6 cells/vial ATCC
xy1231 MN-h 小鼠海马趾神经元 Mouse Neurons-hippocampal 1 x 10^6 cells/vial ATCC
xy1232 MN-sn 小鼠黑质神经元 Mouse Neurons-substantia nigra 1 x 10^6 cells/vial ATCC
xy1233 MN-s 小鼠纹状体神经元 Mouse Neurons-striatal 1 x 10^6 cells/vial ATCC
xy1234 MN-vsc 小鼠腹脊髓神经元 Mouse Neurons-ventral spinal cord 1 x 10^6 cells/vial ATCC
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Cell Death Detection ELISA PLUS roche
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