Whole spinal cord protein lysates from P7 TIMP-1KO and WT mice were separated by SDS-PAGE and Western blotting was performed using anti-GFAP (1:3000; Dako, Denmark), proliferating cell nuclear antigen (PCNA; 1:100, Abcam, Cambridge, MA), and procaspase-3 (1:100; Millipore, Billerica, MA). Equivalent loading was verified by β-actin (1:20 000; Sigma, St Louis, MO). Densitometric measurements of all western blots were performed using ImageJ software (NIH, Bethesda, MD).
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Procaspase-3
Procaspase-3
Procaspase-3 is a key enzyme involved in apoptosis, a form of programmed cell death.
It is the inactive precursor of caspase-3, a critical executioner of apoptosis.
Procaspase-3 plays a central role in the initiation and execution of the apoptotic cascade.
Its activation leads to the cleavage of various cellular substrates, ultimately resulting in cell death.
Understanding the regulation and function of procaspase-3 is crucial for elucidating the mechanisms of apoptosis and its potential therapeutic applications in human health and disease.
Researhers can optimize their procaspase-3 studies using PubCompare.ai, an AI-driven tool that helps identify the most reproducible and accurate findings from the literature, preprints, and patents.
It is the inactive precursor of caspase-3, a critical executioner of apoptosis.
Procaspase-3 plays a central role in the initiation and execution of the apoptotic cascade.
Its activation leads to the cleavage of various cellular substrates, ultimately resulting in cell death.
Understanding the regulation and function of procaspase-3 is crucial for elucidating the mechanisms of apoptosis and its potential therapeutic applications in human health and disease.
Researhers can optimize their procaspase-3 studies using PubCompare.ai, an AI-driven tool that helps identify the most reproducible and accurate findings from the literature, preprints, and patents.
Most cited protocols related to «Procaspase-3»
Actins
Densitometry
Glial Fibrillary Acidic Protein
Mus
Procaspase-3
Proliferating Cell Nuclear Antigen
Proteins
SDS-PAGE
Spinal Cord
Western Blot
Actins
Antibodies
Caspase 3
Cells
Chemiluminescence
Horseradish Peroxidase
Mus
Neurons
Procaspase-3
Proteins
PTEN protein, human
Technique, Dilution
Tissues
VDAC1 protein, human
Western Blot
The anti-Bcl-2 monoclonal antibody was purchased from Dako (Carpinteria, CA, USA). The anti-α-tubulin monoclonal antibody was from Sigma. The anti-poly(ADP-ribosyl)polymerase (PARP) monoclonal, anti-pro-caspase-3 monoclonal, anti-Bcl-xL polyclonal, anti-Mcl-1 polyclonal antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-PKC-α monoclonal antibody was purchased from Neo Transduction Laboratories (Lexington, KY, USA). The antisurvivin polyclonal antibody was purchased from Novus Biological (Littleton, CO, USA). Fetal bovine serum was purchased from Invitrogen. A list of the oligomers employed is presented in Table 1 .
List of the oligomers employed
LNA-modified riboses are given in bold capital letters; whereas small letters indicate deoxyriboses. s, Phosphorothioate; m, C5-methylcytosine; FAM, 5′-fluorescein covalent conjugate.
alpha-Tubulin
bcl-2 Gene
Biopharmaceuticals
Deoxyribose
Fetal Bovine Serum
Fluorescein
Immunoglobulins
Monoclonal Antibodies
Novus
Poly A
PRKCA protein, human
Procaspase-3
Ribose
Cells were grown in 6-well plates for 48 h in hormone-depleted medium and incubated for an additional 1 or 48–72 h with 20–100 μg/mL of Chios mastiha tree polar and medium-polar leaves fractions and/or 10 nM DEX, as indicated. Cells were washed in PBS 1X, lysed in buffer A (20 mM Tris pH:7.5, 250 mM NaCl, 0.5% Triton, 3 mM EDTA) supplemented with cocktail protease inhibitors, DTT and PMSF. After Bradford protein determination, cell extracts were electrophoresed in discontinuous SDS-PAGE and Western blotting with specific antibodies as previously described [48 (link)]. Enhanced chemiluminescence was used for the detection of the protein bands. β-actin and GAPDH expression levels were evaluated for the normalization of the GR, PEPCK, GS, procaspase-3, procaspase-9, Bcl-2, AMPK, and phosphο-AMPKα (Thr172) expression levels. In the case of MG-132 treatment, an inhibitor of the proteasome, HEK293 cells were pre-treated with 5μΜ MG-132, or DMSO, for 1 h [13 (link),14 (link)]. Then, the cell culture medium was replaced and HEK293 cells were further treated with 50 μg/mL leaves fractions or 10 nM DEX or DMSO, for 24 h. Cells were collected, lysed, and subjected to electrophoresis and Western blot analysis.
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Actins
Antibodies
BCL2 protein, human
Buffers
Cell Culture Techniques
Cell Extracts
Cells
Chemiluminescence
Culture Media
Edetic Acid
Electrophoresis
GAPDH protein, human
HEK293 Cells
Hormones
mastiha
MG 132
Phosphoenolpyruvate Carboxylase
Procaspase-3
Procaspase-9
Protease Inhibitors
Proteasome Inhibitor
Proteins
SDS-PAGE
Sodium Chloride
Sulfoxide, Dimethyl
Trees
Tromethamine
Western Blot
Heart tissues were homogenized and extracted using lysis buffer [tissue protein extraction reagent (cat. no. 78510); protease inhibitor cocktail (cat. no. 87785); phosphatase inhibitor cocktail (cat. no. 78420); PMSF (100 µM; cat. no. 36978; all from Thermo Fisher Scientific, Inc.); 100:1:1:1 ratio for configuration], followed by fractionation using a Mitochondrial/Cytosol Fractionation kit (cat. no. ab65320; Abcam) to obtain the cytosolic and mitochondrial fractions. An enhanced BCA protein assay kit (cat. no. P0010; Beyotime Institute of Biotechnology) was used to quantify the protein concentration. The total lysates, cytosolic and mitochondrial fractions (50 µg) were separated via 15% SDS-PAGE and transferred to a nitrocellulose membrane (Immobilon NC; MilliporeSigma). Following blocking in 5% fat-free milk in TBS-0.1% Tween-20 (TBST) for 1 h at room temperature, the membranes were incubated at 4°C overnight with primary antibodies targeted against: CYP2E1 (1:500; cat. no. ab28146), cardiac troponin T (1:500; cTnT; cat. no. ab8295), cytochrome c (1:500; cat. no. ab13575), procaspase 3 (1:1,000; cat. no. ab13847), procaspase 9 (1:1,000; cat. no. ab47537; all from Abcam), cleaved (active) caspase 3 (1:500; cat. no. 9507S) and cleaved caspase 9 (1:500; cat. no. 9664S; both from Cell Signaling Technology, Inc.), anti-β-tubulin (1:1,000; cat. no. ab21058) and anti-voltage dependent anion channel 1 (1:1,000; cat. no. ab14734; both from Abcam) were used. Following washing with TBST, the membranes were incubated with a HRP-conjugated secondary antibody (1:10,000; anti-rabbit IgG, cat. no. ZB-2301 or anti-mouse IgG, cat. no. ZB-2305; ZSGB-BIO, Inc.) at room temperature for 1 h. Primary antibody binding was visualized using a chemiluminescent detection system (Western Blotting Luminal Reagent; Santa Cruz Biotechnology, Inc.) and analyzed using the densitometry function of Quantity One software (version 3.0; Bio-Rad Laboratories, Inc.).
anti-IgG
Antibodies
Biological Assay
Buffers
Cardiac Arrest
Caspase 3
Caspase 9
Cytochrome c1
Cytochrome P-450 CYP2E1
Cytosol
Densitometry
Heart
Immobilon
Immunoglobulins
Milk, Cow's
Mitochondria
Mus
Nitrocellulose
Phenobarbital
Phosphoric Monoester Hydrolases
Procaspase-3
Procaspase-9
Protease Inhibitors
Proteins
Rabbits
Radiotherapy Dose Fractionations
SDS-PAGE
Tissue, Membrane
Tissues
Troponin T
Tubulin
Tween 20
VDAC1 protein, human
Most recents protocols related to «Procaspase-3»
In order to reduce the number of animals used in the present study, 1 mg/kg D. salina was used as the effective dose to investigate the mechanism underlying the cardioprotective effect of D. salina. Protein expression was evaluated by western blot as previously described procedures [44 (link)]. The primary antibodies against COX-2 (Cayman Chemical, Ann Arbor, MI, USA) included TLR4, phospho-JAK2 (p-JAK2), JAK2 (Santa Cruz, Dallas, TX, USA), phospho-NF-κB (p-NF-κB), NF-κB, phospho-IκB (p-IκB), IκB, procaspase-3, cleaved caspase-3 (Cell Signaling, Danvers, MA, USA), signal transducer and activator of transcription-1 (STAT1), phospho-STAT1 (p-STAT1) (Abcam, Cambridge, UK), light chain 3 (LC3), p62 and Beclin-1 (Novus, Centennial, CO, USA). β-actin (Abcam, Cambridge, UK) was used as an internal loading control. The membranes were incubated with an HRP-conjugated secondary antibody (Jackson Immuno Research Laboratories, West Grove, PA, USA) prior to chemiluminescence detection (Thermo Scientific, Waltham, MA, USA).
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Actins
Animals
Antibodies
BECN1 protein, human
Caimans
Caspase 3
Chemiluminescence
Immunoglobulins
JAK2 protein, human
Light
Novus
Procaspase-3
Proteins
PTGS2 protein, human
RELA protein, human
Tissue, Membrane
Transcription, Genetic
Transducers
Western Blot
The SVT2 were seeded on six-well plates at a density of 2.5 × 105 cells/well, whereas A431 was seeded at a density of 3 × 105 cells/well. After 24 h, cells were treated with 9.8 μM of 1-Me,Me or 2.3 μM 1-Oct,Me . After 48 h of incubation, Western blot analyses were performed by using pro-caspase 9 (Cell Signaling Technology, Danvers, MA, USA) and pro-caspase 3 (Abcam, Cambridge, MA, USA) antibodies, as reported by Del Giudice et al. [48 (link)] Protein intensity levels were normalized using β-actin (Sigma-Aldrich, St. Louis, MO, USA). The chemiluminescence detection system was purchased from Bio-Rad (Hercules, CA, USA).
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Actins
Antibodies
Cells
Chemiluminescence
link protein
Procaspase-3
Procaspase-9
Western Blot
Caspase-Glo 3/7 Assay (Promega, Madison, WI, USA) was performed as per the manufacturer’s protocol. At the indicated time points, mock and ZIKV infected FRPE and iRSCs were incubated with the pro-luminescent caspase-3/7 substrate for 1 h at room temperature. Subsequently, 100 µL of lysate was transferred to a white 96-well microtiter plate for reading the luminescence signal using a luminometer (Glomax Microplate Luminometer, Promega, Madison, WI, USA).
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Biological Assay
Caspase-7
Luminescence
Procaspase-3
Promega
Zika Virus
An equal amount of proteins from whole-cell lysates were separated by 4–12% SDS-PAGE and were transferred to a nitrocellulose membrane. Blots were blocked for 1 h with 5% non-fat dry milk and then incubated over night with the following primary antibodies: anti-CA-IX (R&D), anti-N-cadherin, anti-β-catenin and anti-Vimentin (CST-9782), anti-pro-Caspase-3, anti-Cleaved-Caspase-3, anti-AKT (CST-9272), anti-pAKT (CST-9271), anti-ERK (CST-9102), anti-pERK (CST-9101), anti-MMP-2 (CST-33437), anti-CD44 (Abcam), anti-GADPH (G8795) and anti-Actin (A4700). After washing with 0.1% Tween-20 in PBS, the filters were incubated with their respective secondary antibodies for 1 h and analyzed using the ECL system. Densitometric analyses were performed on at least two different expositions to assure the linearity of each acquisition using ImageJ software (v1.46r) [23 (link)].
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Actins
Antibodies
CA9 protein, human
Cadherins
Caspase 3
CD44 protein, human
Cells
CTNNB1 protein, human
Densitometry
Milk, Cow's
MMP2 protein, human
Nitrocellulose
Procaspase-3
Proteins
SDS-PAGE
Tissue, Membrane
Tween 20
Vimentin
The level of procaspase-3 was quantified using a colorimetric protease assay kit from Invitrogen (Waltham, MA, USA). The level of cleaved caspase-3, a key mediator of apoptosis, was assessed using an ELISA kit from R&D Systems (Minneapolis, MN, USA).
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Apoptosis
Biological Assay
Caspase 3
Colorimetry
Endopeptidases
Enzyme-Linked Immunosorbent Assay
Procaspase-3
Top products related to «Procaspase-3»
Sourced in United States, United Kingdom, China
Procaspase-3 is a laboratory reagent used for the detection and measurement of caspase-3 activity. Caspase-3 is a critical executioner of apoptosis, or programmed cell death, and plays a central role in the apoptotic process.
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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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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.
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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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Procaspase-9 is a laboratory reagent used in research studies. It is an inactive precursor form of the caspase-9 enzyme, which plays a central role in the initiation of the apoptotic (programmed cell death) signaling pathway.
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β-actin is a cytoskeletal protein that is ubiquitously expressed in eukaryotic cells. It is an important component of the microfilament system and is involved in various cellular processes such as cell motility, structure, and integrity.
Sourced in United States
Procaspase-3 is a key enzyme involved in the initiation of apoptosis, or programmed cell death. It serves as a precursor to the active caspase-3 enzyme, which plays a central role in the execution phase of apoptosis. The Procaspase-3 product can be used for research purposes to study the mechanisms of cell death and apoptosis.
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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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β-actin is a cytoskeletal protein that is ubiquitously expressed in eukaryotic cells. It is a component of the microfilament system and plays a crucial role in various cellular processes, such as cell motility, maintenance of cell shape, and intracellular trafficking.
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β-actin is a protein that is found in all eukaryotic cells and is involved in the structure and function of the cytoskeleton. It is a key component of the actin filaments that make up the cytoskeleton and plays a critical role in cell motility, cell division, and other cellular processes.
More about "Procaspase-3"
Procaspase-3 is a crucial enzyme involved in apoptosis, a form of programmed cell death.
It is the inactive precursor of caspase-3, a critical executioner of this cellular self-destruction process.
Procaspase-3 plays a central role in the initiation and execution of the apoptotic cascade.
Its activation leads to the cleavage of various cellular substrates, ultimately resulting in cell death.
Understanding the regulation and function of this proenzyme is crucial for elucidating the mechanisms of apoptosis and its potential therapeutic applications in human health and disease.
Researchers can optimize their Procaspase-3 studies using PubCompare.ai, an AI-driven tool that helps identify the most reproducible and accuarate findings from the literature, preprints, and patents.
This innovative solution allows you to compare multiple methods side-by-side, streamlining your research process and helping you obtain reliable results.
Key related terms and concepts include Cleaved caspase-3 (the active form of the enzyme), PVDF membranes (used for Western blotting to detect Procaspase-3 and Cleaved caspase-3), FBS (fetal bovine serum, a common cell culture supplement), Procaspase-9 (another crucial apoptosis-related enzyme), β-actin (a common loading control protein), and Bcl-2 (a protein that regulates apoptosis).
By understanding these interconnected topics, researchers can design more comprehensive and effective Procaspase-3 studies.
It is the inactive precursor of caspase-3, a critical executioner of this cellular self-destruction process.
Procaspase-3 plays a central role in the initiation and execution of the apoptotic cascade.
Its activation leads to the cleavage of various cellular substrates, ultimately resulting in cell death.
Understanding the regulation and function of this proenzyme is crucial for elucidating the mechanisms of apoptosis and its potential therapeutic applications in human health and disease.
Researchers can optimize their Procaspase-3 studies using PubCompare.ai, an AI-driven tool that helps identify the most reproducible and accuarate findings from the literature, preprints, and patents.
This innovative solution allows you to compare multiple methods side-by-side, streamlining your research process and helping you obtain reliable results.
Key related terms and concepts include Cleaved caspase-3 (the active form of the enzyme), PVDF membranes (used for Western blotting to detect Procaspase-3 and Cleaved caspase-3), FBS (fetal bovine serum, a common cell culture supplement), Procaspase-9 (another crucial apoptosis-related enzyme), β-actin (a common loading control protein), and Bcl-2 (a protein that regulates apoptosis).
By understanding these interconnected topics, researchers can design more comprehensive and effective Procaspase-3 studies.