Immunofluorescence analysis was performed as previously described 30 (link). Briefly, NP cells or tissues attached to slides were fixed with 4% paraformaldehyde, then permeabilized with 0.2% Triton X-100 in PBS. The slides were washed in PBS and blocked with 2% bovine serum albumin (BSA) in PBS for 2 h at 37 °C, and then incubated with primary antibodies against: GRP78 (1:50), GRP94 (1:50), CHOP (1:100), caspase-3 (1:50), or caspase-12 (1:100) (Proteintech). After washed twice, the slides were subsequently treated with secondary goat anti-rabbit antibody (Boster) at 37 °C for 2 h. Nuclei were co-stained for 5 min with 0.1 g/mL DAPI (Beyotime, Nantong, China), and images were captured under a microscope (Olympus, BX53; Melville, NY, USA).
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Caspase 12
Caspase 12
Caspase 12 is a member of the cysteine-aspartic acid protease (caspase) family, which play a central role in apoptosis, or programmed cell death.
This enzyme is primarily expressed in the endoplasmic reticulum and is thought to be involved in the cellular response to endoplasmic reticulum stress.
Caspase 12 has been implicated in neurodegenerative diseases and may also play a role in the immune response.
Researching the functions and regulation of this important protein can provide valuable insights into cellular processes and disease pathogenesis.
Discover the power of Caspase 12 research optimization with PubCompare.ai's AI-driven platform.
Unlcok access to protocols from literature, pre-prints, and patents, and leverage intelligent comparisons to identify the best strategies and products for your research.
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This enzyme is primarily expressed in the endoplasmic reticulum and is thought to be involved in the cellular response to endoplasmic reticulum stress.
Caspase 12 has been implicated in neurodegenerative diseases and may also play a role in the immune response.
Researching the functions and regulation of this important protein can provide valuable insights into cellular processes and disease pathogenesis.
Discover the power of Caspase 12 research optimization with PubCompare.ai's AI-driven platform.
Unlcok access to protocols from literature, pre-prints, and patents, and leverage intelligent comparisons to identify the best strategies and products for your research.
Unleash your potentinal with PubCompare.ai's cutting-edge tools.
Most cited protocols related to «Caspase 12»
Antibodies
Antibodies, Anti-Idiotypic
Caspase 3
Caspase 12
Cell Nucleus
Cells
DAPI
DDIT3 protein, human
Fluorescent Antibody Technique
Glucose Regulated Protein 78 kDa
Goat
GRP94
Microscopy
paraform
Rabbits
Serum Albumin, Bovine
Tissues
Triton X-100
Antibodies
ATF4 protein, human
Brain
Calnexin
Caspase 3
Caspase 9
Caspase 12
Cathepsin D
Chemiluminescence
Cytochromes c'
DDIT3 protein, human
Densitometry
Dot Immunoblotting
Glucose Regulated Protein 78 kDa
GRP94
Homo sapiens
Horseradish
Immunoblotting
Immunoglobulins
Mus
Nitrocellulose
Oncogenes
Proteins
Synaptotagmins
Syntaxin 6
TCL1B protein, human
Tissue, Membrane
Tissues
Ubiquitin
X-Ray Film
Apoptotic and necrotic β-cells and INS-1E cells were counted in fluorescence microscopy following staining with the DNA-binding dyes propidium iodide (5 μg/ml) and Hoechst 33342 (10 μg/ml) (11 (link)). The percentage human islet cell death was determined as described (12 (link)). The apoptosis index was calculated as [(% apoptosis in ER stressor–treated cells under indicated conditions transfected with negative or target siRNA − % apoptosis in ER stressor–treated negative siRNA-transfected cells)/(100% − % apoptosis in ER stressor–treated negative siRNA-transfected cells)] × 100. Apoptosis was also assessed by the Cell Death Detection ELISAplus kit (Roche Diagnostics, Mannheim, Germany) (10 (link)). Forskolin did not modify INS-1E cell proliferation under the present experimental conditions (assessed by BrdU incorporation, data not shown).
To study cytochrome C release, cells were prepared as described in the online appendix supplementary Methods. After total protein quantification (Bio-Rad Laboratories, Munchen, Germany), equal amounts of protein were blotted using tubulin as cytoplasmic and apoptosis-inducing factor (AIF) as mitochondrial control.
Caspase 3 and 7 activation was measured with the Caspase-Glo 3/7 assay (Promega, Madison, WI) and caspase 12 activation with the CaspGlow Caspase-12 staining kit (Biovision, Mountain View, CA) (12 (link)).
To study cytochrome C release, cells were prepared as described in the online appendix supplementary Methods. After total protein quantification (Bio-Rad Laboratories, Munchen, Germany), equal amounts of protein were blotted using tubulin as cytoplasmic and apoptosis-inducing factor (AIF) as mitochondrial control.
Caspase 3 and 7 activation was measured with the Caspase-Glo 3/7 assay (Promega, Madison, WI) and caspase 12 activation with the CaspGlow Caspase-12 staining kit (Biovision, Mountain View, CA) (12 (link)).
Apoptosis
Apoptosis Inducing Factor
Biological Assay
Bromodeoxyuridine
Caspase-7
Caspase 3
Caspase 12
Cell Death
Cell Proliferation
Cells
Colforsin
Cytochromes c'
Cytoplasm
Diagnosis
Dyes
HOE 33342
Homo sapiens
Islets of Langerhans
Microscopy, Fluorescence
Necrosis
Promega
Propidium Iodide
Proteins
RNA, Small Interfering
Tubulin
Tissues or cultured cells were rinsed in ice-cold PBS and were immediately homogenized in 5 vol Triton X-100 buffer (20 mM Hepes, pH 7.5, 150 mM NaCl, 1% Triton X-100, 10% glycerol, 1 mM EDTA, 10 mM tetrasodium pyrophosphate, 100 mM NaF, 17.5 mM β-glycerophosphate, 10 mM PMSF, 15 μg/ml aprotonin, and 6 μg/ml pepstatin A) using a motorized homogenizer. After incubating on ice for 15 min, the extracts were cleared by centrifugation at 14,000 rpm twice for 30 min each. The protein content of each extract was determined by protein assay (Bio-Rad Laboratories). The extracts (40 μg) were separated by SDS-PAGE and were transferred to nitrocellulose. The blots were incubated with primary antibody (see below), and the signal was revealed by chemiluminescence after reacting with HRP-conjugated second antibody. The following primary antibodies were used: anti–eIF-2α (1:500; Santa Cruz Biotechnology, Inc.); anti–p-eIF-2α (1:1,000; Cell Signaling Technology); anti–caspase-12 (1:500; Santa Cruz Biotechnology, Inc.); and antiactin (1:1,000; Sigma-Aldrich).
Antibodies
beta-glycerol phosphate
Biological Assay
Buffers
Caspase 12
Centrifugation
Chemiluminescence
Cold Temperature
Cultured Cells
Edetic Acid
Glycerin
HEPES
Immunoglobulins
Nitrocellulose
pepstatin
Proteins
SDS-PAGE
Sodium Chloride
sodium pyrophosphate
Tissues
Triton X-100
The liver tissue was homogenized as described previously.31 (link), 32 (link) Total liver lysates were then used to quantify GRP78, CHOP, total PERK and pPERK, total and peIF2α, ATF4, ATF6α, ATF6β, sXBP-1, TRAF2, cleaved caspase 3, caspase 12, cytochrome c, total and phospho-ERK, total and phospho-p38 MAPK, total and phospho-GSK3β, total and phospho-JNK, total and phospho-AKT, as well as total and phospho-VDAC by western blot. Cytosolic fractions were used to quantify cleaved caspase 9 and cytochrome c by western blot. Proteins were separated by SDS-PAGE and transferred into polyvinylidene fluoride membranes. Membranes were immunoblotted with antibodies directed against GRP78, CHOP, total and pPERK, total and peIF2α, ATF4, ATF6α, ATF6β, sXBP-1, β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and TRAF2, cleaved caspase 3, cleaved 9 and caspase 12, cytochrome c, total and phospho-ERK, total and phospho-p38 MAPK, total and phospho-GSK3β, total and phospho-JNK and total and phospho-AKT (Cell Signalling Technology Inc., Beverly, MA, USA). The bands were visualized using an enhanced chemiluminescence kit (Bio-Rad Laboratories, Hercules, CA, USA). The values were obtained by densitometric scanning and the Quantity One software program (Bio-Rad Laboratories, Hercules, CA, USA). The scanning values for GRP78, CHOP, ATF4, ATF6α, ATF6β, sXBP-1, TRAF2, cleaved caspase 3, cleaved caspase 9, caspase 12 and cytochrome c were divided by the scanning values for β-actin, and those for phosphorylated PERK, eIF2α, GSK3β, VDAC, ERK, JNK and p38 MAPK by the total PERK, eIF2α, GSK3β, VDAC, ERK, JNK and p38 MAPK, respectively.
Actins
Antibodies
ATF4 protein, human
Caspase 3
Caspase 9
Caspase 12
Chemiluminescence
Cytochromes c
Cytosol
DDIT3 protein, human
Glucose Regulated Protein 78 kDa
GSK3B protein, human
Liver
Mitogen-Activated Protein Kinase p38
polyvinylidene fluoride
Proteins
SDS-PAGE
Tissue, Membrane
Tissues
TNF Receptor Associated Factor 2
Western Blot
Most recents protocols related to «Caspase 12»
Anti-PERK (3192), anti-p-PERK (3179S), anti-CHOP (2895), anti-caspase-4 (4450), anti-cleaved caspase-3 (Cleaved caspase-3) (9664) and anti-GAPDH (5174) antibodies were obtained from Cell Signaling (Danvers, USA). Anti-CHOP (15204-1-AP), anti-78-kDa glucose-regulated protein (GRP78) (66574-1-Ig), anti-Nuclear respiratory factor 2 (Nrf2) (16396-1-AP), anti-Keap1 (60027-1-Ig), anti-GAPDH (10494-1-AP) and anti-β-actin (20536-1-AP) were obtained from Proteintech (Chicago, USA). Anti-apelin (ab125213) and anti-caspase-12 (ab62484) were obtained from Abcam (Cambridge, UK). Anti-β-tubulin (GB11017) was obtained from Servicebio (Wuhan, China). All secondary antibodies were obtained from Abcam (Cambridge, UK). Apelin-13 (A6469, purity ≥ 95.0%) and 4-phenylbutyrate (4-PBA) (P21005) were purchased from Sigma-Aldrich (St. Louis, USA). GSK2656157 (5.04651) was purchased from EMD Millipore (Massachusetts, USA). Tunicamycin (TM) (ab120296) was purchased from Abcam (Cambridge, UK). Iohexol was purchased from GE Healthcare (Shanghai, China).
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4-PBA
4-phenylbutyrate
Actins
Antibodies
apelin-13 peptide
APLN protein, human
CASP4 protein, human
Caspase 3
Caspase 12
DDIT3 protein, human
GA-Binding Protein Transcription Factor
GAPDH protein, human
Glucose Regulated Protein 78 kDa
GSK2656157
Iohexol
KEAP1 protein, human
Tubulin
Tunicamycin
Anti-PERK (3192), anti-p-PERK (3179S), anti-CHOP (2895), anti-caspase-4 (4450), anti-cleaved caspase-3 (Cleaved caspase-3) (9664) and anti-GAPDH (5174) antibodies were obtained from Cell Signaling (Danvers, USA). Anti-CHOP (15204-1-AP), anti-78-kDa glucose-regulated protein (GRP78) (66574-1-Ig), anti-Nuclear respiratory factor 2 (Nrf2) (16396-1-AP), anti-Keap1 (60027-1-Ig), anti-GAPDH (10494-1-AP) and anti-β-actin (20536-1-AP) were obtained from Proteintech (Chicago, USA). Anti-apelin (ab125213) and anti-caspase-12 (ab62484) were obtained from Abcam (Cambridge, UK). Anti-β-tubulin (GB11017) was obtained from Servicebio (Wuhan, China). All secondary antibodies were obtained from Abcam (Cambridge, UK). Apelin-13 (A6469, purity ≥ 95.0%) and 4-phenylbutyrate (4-PBA) (P21005) were purchased from Sigma-Aldrich (St. Louis, USA). GSK2656157 (5.04651) was purchased from EMD Millipore (Massachusetts, USA). Tunicamycin (TM) (ab120296) was purchased from Abcam (Cambridge, UK). Iohexol was purchased from GE Healthcare (Shanghai, China).
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4-PBA
4-phenylbutyrate
Actins
Antibodies
apelin-13 peptide
APLN protein, human
CASP4 protein, human
Caspase 3
Caspase 12
DDIT3 protein, human
GA-Binding Protein Transcription Factor
GAPDH protein, human
Glucose Regulated Protein 78 kDa
GSK2656157
Iohexol
KEAP1 protein, human
Tubulin
Tunicamycin
For immunoprecipitation, cells were lysed with NP40 lysis buffer (50 mM Tris-HCl pH 7.5, 100 mM NaCl, 1% NP-40, 1 mM EDTA, 1 mM DTT, 10% glycerol) containing protease inhibitors. After centrifugation, the supernatants were incubated with antibody overnight and then Protein A/G agarose for 2 h at 4 °C. Immunocomplexes were washed and analyzed by Western blot. For Western blot, the proteins from lysed cells were denatured and separated with SDS-PAGE. Then, the proteins were transferred to PVDF membranes, blocked and incubated with the corresponding primary and secondary antibodies. The specific bands were analyzed by the Western blot infrared imaging system (LI-COR Biosciences). The following antibodies were used: EVA1A: A8070, ABclonal; CHOP: 2895, Cell signaling Technology (CST); IRE1: 14C10, CST; ATF6: 65880 T, CST; Actin: AC026, ABclonal; Flag: 20543-1-AP, Proteintech; Caspase 8: 4927, CST; C-Caspase 9: 9509, CST; C-Caspase 3: 9664, CST; Caspase 12: 2202, CST; C-Parp: 9548, CST; GFP: ab290, Abcam; MCL1: 94296, CST; Bak: 12105, CST; LC3: 27543,Sigma; P62: 18420-1-AP, Proteintech; BNIP3: ab10433, Abcam; TIM23: 111263-A, Proteintech; TOM20: ab56783, Abcam.
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Actins
activating transcription factor 6, human
Antibodies
Buffers
Caspase-8
Caspase 3
Caspase 9
Caspase 12
Centrifugation
DDIT3 protein, human
Edetic Acid
G-substrate
Glycerin
GTP-Binding Proteins
Immunoglobulins
Immunoprecipitation
MCL1 protein, human
Nonidet P-40
polyvinylidene fluoride
Protease Inhibitors
Proteins
SDS-PAGE
Sepharose
Sodium Chloride
Staphylococcal Protein A
Tissue, Membrane
Tromethamine
Western Blot
Total protein was extracted using RIPA kit (AS1004, ASPEN), and protease inhibitors (AS1008, ASPEN) were added during protein extraction to prevent protein degradation. After electrophoresis, coating and sealing, the indexes to be detected were incubated with primary antibody and secondary antibody, and finally immunolabeled with enhanced chemiluminescence reagent (AS1059, ASPEN). Antibodies against caspase-3 (#9662), caspase-12 (#9671), phosphorylated PERK (p-PERK, Thr980, #3179), PERK (#3192), phosphorylated eIF2α (p-eIF2α, Ser51, #3597), eIF2α (#2103), GRP78 (#3183), CHOP (#2895), and HO-1 (#43966) were obtained from cell signing technology corporation (Massachusetts, USA). Anti-β-tubulin (#ab6046), anti-β-actin (#ab8226), and anti-Nrf-2 (#ab137550) were purchased from Abcam (Cambridge, USA). Antihistone H3 (EM1108) was purchased from ELK Biotechnology (Wuhan, China).
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Actins
Antibodies
Caspase 3
Caspase 12
Cells
Chemiluminescence
DDIT3 protein, human
Electrophoresis
GA-Binding Protein Transcription Factor
Glucose Regulated Protein 78 kDa
Immunoglobulins
Protease Inhibitors
Proteins
Proteolysis
Radioimmunoprecipitation Assay
Tubulin
Antibodies against the following proteins were used in this study: Bip (3177 s, Cell Signaling Technology [CST]), eIF2α (5324P, CST), Phospho-eIF2α (3398P, CST), SAPK/JNK (9252S, CST), Phospho-SAPK/JNK (9255S, CST), Caspase-12(2202S, CST), Cleaved Caspase-3 (9661S, CST), Cleaved PARP (9541S, CST), Phospho-EGFR and EGFR (11862S, CST), GAPDH (60,004–1-Ig, Proteintech). PE anti-human EGFR Antibody (352,904, Biolegend). The kinase inhibitor library was purchased from Shanghai Topscience Co., Ltd (Shanghai, China). Afatinib (HY-10261) were purchased from MCE.
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Afatinib
Antibodies
Antibodies, Anti-Idiotypic
CASP3 protein, human
Caspase 12
cDNA Library
EGFR protein, human
GAPDH protein, human
Homo sapiens
Phosphotransferases
Proteins
Top products related to «Caspase 12»
Sourced in United States
Caspase-12 is a laboratory tool used to detect and quantify the presence of the Caspase-12 protein in biological samples. Caspase-12 is an enzyme involved in the process of apoptosis, or programmed cell death. This product can be utilized in various research applications that require the measurement or analysis of Caspase-12 levels.
Sourced in United States, United Kingdom, China
Caspase-12 is an enzyme involved in the process of programmed cell death, known as apoptosis. It plays a role in the activation of the apoptotic pathway in response to certain cellular stresses. Caspase-12 is primarily found in the endoplasmic reticulum and is considered a key player in the endoplasmic reticulum stress-induced apoptotic pathway.
Sourced in United States
Anti-caspase-12 is a primary antibody that specifically binds to and detects caspase-12, a member of the caspase family of proteases. Caspase-12 is involved in apoptosis, or programmed cell death, and is implicated in various cellular processes.
Sourced in United States, Germany, China, United Kingdom, Morocco, Ireland, France, Italy, Japan, Canada, Spain, Switzerland, New Zealand, India, Hong Kong, Sao Tome and Principe, Sweden, Netherlands, Australia, Belgium, Austria
PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
Sourced in United States, United Kingdom
Ab62484 is a rabbit polyclonal antibody that recognizes the CD44 antigen. CD44 is a cell-surface glycoprotein involved in cell-cell interactions, cell adhesion and migration.
Sourced in United States, United Kingdom, China, Germany, Canada, Japan, Morocco, Denmark, Switzerland, France, Netherlands, Macao
Cleaved caspase-3 is an antibody that detects the activated form of caspase-3 protein. Caspase-3 is a key enzyme involved in the execution phase of apoptosis, or programmed cell death. The cleaved caspase-3 antibody specifically recognizes the active, cleaved form of the enzyme and can be used to monitor and quantify apoptosis in experimental systems.
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Bcl-2 is a protein that plays a key role in regulating apoptosis, or programmed cell death. It functions as an anti-apoptotic protein, helping to prevent cell death by inhibiting the activity of pro-apoptotic proteins.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Caspase-3 is a key enzyme involved in the execution phase of cell apoptosis (programmed cell death). It plays a central role in the apoptotic pathway by cleaving various cellular substrates, leading to the characteristic morphological and biochemical changes associated with apoptosis.
Sourced in United States, China, United Kingdom
GRP78 is a molecular chaperone protein that plays a crucial role in the endoplasmic reticulum (ER) of eukaryotic cells. It is involved in the folding and assembly of proteins within the ER, helping to maintain ER homeostasis and prevent the accumulation of misfolded proteins.
More about "Caspase 12"
Caspase-12, a member of the cysteine-aspartic acid protease (caspase) family, plays a central role in apoptosis, or programmed cell death.
This enzyme is primarily expressed in the endoplasmic reticulum (ER) and is thought to be involved in the cellular response to ER stress.
Caspase-12 has been implicated in neurodegenerative diseases and may also play a role in the immune response.
Researching the functions and regulation of this important protein can provide valuable insights into cellular processes and disease pathogenesis.
Caspase-12 is a key regulator of the apoptotic pathway, and its dysregulation has been linked to a variety of disease states, including Alzheimer's disease, Parkinson's disease, and ischemic brain injury.
To optimize your Caspase-12 research, explore the AI-driven platform at PubCompare.ai.
Unlock access to protocols from literature, pre-prints, and patents, and leverage intelligent comparisons to identify the best strategies and products for your research.
Discover the power of PubCompare.ai's cutting-edge tools and unleash your potential in understanding the role of Caspase-12 in cellular processes and disease pathogenesis.
Delve deeper into the world of Caspase-12 by exploring related topics such as Anti-caspase-12, PVDF membranes, Ab62484, Cleaved caspase-3, Bcl-2, FBS, and Caspase-3.
Gain a holistic understanding of the intricate signaling pathways and regulatory mechanisms involving Caspase-12 and its interacting partners.
With the right tools and resources, you can unlock new avenues for Caspase-12 research and drive advancements in our understanding of cellular processes and disease pathogenesis.
This enzyme is primarily expressed in the endoplasmic reticulum (ER) and is thought to be involved in the cellular response to ER stress.
Caspase-12 has been implicated in neurodegenerative diseases and may also play a role in the immune response.
Researching the functions and regulation of this important protein can provide valuable insights into cellular processes and disease pathogenesis.
Caspase-12 is a key regulator of the apoptotic pathway, and its dysregulation has been linked to a variety of disease states, including Alzheimer's disease, Parkinson's disease, and ischemic brain injury.
To optimize your Caspase-12 research, explore the AI-driven platform at PubCompare.ai.
Unlock access to protocols from literature, pre-prints, and patents, and leverage intelligent comparisons to identify the best strategies and products for your research.
Discover the power of PubCompare.ai's cutting-edge tools and unleash your potential in understanding the role of Caspase-12 in cellular processes and disease pathogenesis.
Delve deeper into the world of Caspase-12 by exploring related topics such as Anti-caspase-12, PVDF membranes, Ab62484, Cleaved caspase-3, Bcl-2, FBS, and Caspase-3.
Gain a holistic understanding of the intricate signaling pathways and regulatory mechanisms involving Caspase-12 and its interacting partners.
With the right tools and resources, you can unlock new avenues for Caspase-12 research and drive advancements in our understanding of cellular processes and disease pathogenesis.