Capsaicin (CAP) and Sorafenib were purchased to Sigma (St. Louis, MO, USA). Primary antibodies anti-caspase-9, anti-PARP, anti-AFP, anti-pAkt-ser473, p-mTOR-ser2448, p-AMPKα1-thr172, p-ACC-ser79 and the antibodies against the corresponding total forms were obtained from Cell Signaling Technology (Danvers, MA, USA). The anti-caspase-3 antibody was from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). (Peroxidase labeled secondary anti-mouse IgG was from Sigma (St. Louis, MO, USA) and anti-rabbit IgG was from Calbiochem (San Diego, CA, USA).
Anti caspase 3 antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Anti-caspase-3 antibody is a laboratory reagent used to detect and measure the expression of caspase-3, a key enzyme involved in the apoptosis (programmed cell death) pathway. This antibody can be used in various immunoassay techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the role of caspase-3 in biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti bcl 2 antibody, by Santa Cruz Biotechnology (2 mentions)
Anti vegf antibody, by Santa Cruz Biotechnology (2 mentions)
Anti β actin antibody, by Santa Cruz Biotechnology (2 mentions)
Lsm 510, by Zeiss (2 mentions)
Anti bax antibody, by Santa Cruz Biotechnology (2 mentions)
Anti β actin, by Abcam (2 mentions)
Sorafenib, by Merck Group (1 mentions)
P mtor ser2448, by Cell Signaling Technology (1 mentions)
Peroxidase labeled secondary anti mouse igg, by Merck Group (1 mentions)
P ampkα1 thr172, by Cell Signaling Technology (1 mentions)
P acc ser79, by Cell Signaling Technology (1 mentions)
Capsaicin cap, by Merck Group (1 mentions)
Anti afp, by Cell Signaling Technology (1 mentions)
Anti p akt ser473, by Cell Signaling Technology (1 mentions)
Anti caspase 9, by Cell Signaling Technology (1 mentions)
Anti parp, by Cell Signaling Technology (1 mentions)
Anti n smase, by Abcam (1 mentions)
β actin, by Abcam (1 mentions)
Anti c myc antibody, by Santa Cruz Biotechnology (1 mentions)
Sc 7480, by Santa Cruz Biotechnology (1 mentions)
21 protocols using anti caspase 3 antibody
1
Apoptosis and Proliferation Signaling
2
Caspase-3 Immunofluorescence in H9c2 Cells
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H9c2 cells were fixed in 4% paraformaldehyde for 15 min at RT and permeabilized with 0.5% TritonX-100 for 5 min. After permeabilization, the cells were blocked by incubation with 10% FBS at RT for 1 h. Then, H9c2 cells were incubated with anti-Caspase-3 antibody (1:50, Santa Cruz) at 4 °C overnight. Next, the cells were washed three times with PBS and incubated with secondary anti-mouse FITC (diluted 1:100) in the dark at RT for 1 h. Nuclei were stained for 10 min with DAPI. After three times washes in PBS, coverslips were mounted on slides with anti-fade mounting medium and evaluated under a laser confocal microscope (Carl Zeiss, LSM 510).
3
Western Blot Analysis of Apoptosis Markers
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Tissues dissected from the mice were homogenized. The western blot analysis was performed based on the protocol described in our previous study25 (link). The optical density of each band was measured using Image J. Anti-SPT-1 (1:300), anti-N-SMase (1:1,000), and β-actin (1:5,000) antibodies were procured from Abcam (Cambridge, MA, USA). Anti-acidic SMase (A-SMase) antibody (1: 500) and anti-caspase-3 antibody were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and Cell signaling Technology (Danvers, MA, USA), respectively.
4
Molecular Mechanisms of Cardiac Remodeling
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The experimental protocol was approved by the Animal Care and Use Committee of Xuzhou Medical College. Rats were housed in a climate-controlled room. Sterile water and standard chow diet were available ad libitum. Six-week-old male Sprague-Dawley (SD) rats were provided by the Animal Department, Xuzhou Medical College. All of the antibodies, including anti-forkhead box H1 (Foxh1) antibody, anti-c-Myc antibody, anti-p-Smad3 antibody, anti-matrix metalloproteinase 2 (MMP2) antibody, anti-tissue inhibitor of matrix metalloproteinases type 2 (TIMP2) antibody, anti-Bax antibody, anti-Bcl-2 antibody, anti- Caspase-3 antibody and anti-APJ antibody, were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All other reagent-grade chemicals were purchased from the Sigma- Aldrich Chemical Co. (St. Louis, MO, USA).
5
Western Blot Analysis of Cell Signaling
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The cell lysates (20 µg) underwent electrophoresis and were transferred to the nitrocellulose membrane. The latter were blocked in 5% of non-fat milk in PBS 0.1% Tween-20, then incubated in the presence of mouse monoclonal anti-β actin antibody (antibody dilution 1:5000) (A5316 Sigma, St. Loius, MO, USA), rabbit polyclonal anti-Nitric Oxide Synthase-2 (NOS-2) antibodies (antibody dilution 1:200) (sc-651 purchased by Santa Cruz biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-Matrix Metalloproteinases-9 (MMP-9), and anti-Bax antibodies (antibodies dilution 1:200) (sc-5302 and sc-7480, respectively, both purchased by Santa Cruz biotechnology, Santa Cruz, CA, USA), goat polyclonal anti-caspase-3 antibody (antibody dilution 1:200) (sc-1225 purchased by Santa Cruz biotechnology, Santa Cruz, CA, USA). Samples were then probed with specific enzyme conjugated IgG horseradish peroxidase. Immunoreactive bands were revealed by ECL system (Amersham Int., Buckunghamshire, UK) and underwent densitometric analysis. Values obtained from densitometry, expressed as Integrated Optical Intensity (IOI), were evaluated with a CHEMIDOC XRS system through the QuantiOne 1-D analysis software (BIORAD, Richmond, CA, USA). Data were normalized with densitometric values derived from β-actin loading control.
6
Protein Expression Analysis by Western Blot
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The protein expression levels of MDM2, p53, VEGF, caspase-3, cleaved caspase-3, and poly(ADP-ribose) polymerase (PARP) were analyzed by western blot. The following antibodies were used: anti-MDM2 antibody (1:1000; Sigma), anti-p53 antibody (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-VEGF antibody (1:200; Santa Cruz Biotechnology), anti-caspase-3 antibody (1:1000; Santa Cruz Biotechnology), anti-cleaved caspase-3 p11 antibody (1:1000; Santa Cruz Biotechnology), and anti-PARP antibody (1:1000; Cell Signaling, Danvers, MA, USA).
7
Antibody Optimization for Western Blot
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Primary antibodies for Western blot analysis were as follows: anti-LOX1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA), anti-STRA6 antibody (ABGENT, San Diego, CA), anti-CRBP1 antibody (Santa Cruz Biotechnology), anti-RARα antibody (Santa Cruz Biotechnology), anti-RARγ antibody (Santa Cruz Biotechnology), anti-RXRα antibody (Santa Cruz Biotechnology), anti-c-Jun N-terminal kinase (JNK) antibody (Santa Cruz Biotechnology), anti-pJNK antibody (Abcam, Cambridge, MA), anti-p38MAPK antibody (ABGENT), anti-p-p38MAPK antibody (ABGENT), anti-pSmad2 antibody (Santa Cruz Biotechnology), anti-Smad2 antibody (Santa Cruz Biotechnology), anti-TGFβ1 (Santa Cruz Biotechnology), anti-caspase 3 antibody (Santa Cruz Biotechnology), anti-collagen 1 antibody (Santa Cruz Biotechnology), and anti-actin antibody (Millipore, Temecula, CA). Secondary antibodies for Western blot analysis as HRP-conjugate antibody were purchased from Millipore. The inhibitors of JNK, SP600125 and p38MAPK, SB203580 were purchased from Sigma-Aldrich (St. Louis, MO).
8
Apoptosis Assay Protocol for Cell Lines
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Calcium ionophore (A23187), benzamidine hydrochloride, N-acetyl-Leu-Glu-His-Asp trifluoro methylcoumarin (AC-LEHD-FMC), acetyl-Asp-Glu-Val-Asp-7-amido-4-methylcoumarin (AC-DEVD-AMC), sodium orthovanadate, dimethyl sulfoxide (DMSO), fluorescein isothiocyanate (FITC)-labeled annexin V, 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate acetylester (CM-H2DCFDA), CHAPS, rhodamine 123, leupeptin hydrochoride, N-(2-Hydroxyethyl) piperazine-N′-ethanesulfonic acid (HEPES), fura-2/AM, monoclonal anti-phosphotyrosine antibody, acridine orange 10-nonyl bromide (NAO) and dithiothreitol (DTT) were from Sigma Chemicals, St. Louis (USA). Monoclonal anti-cytochrome c antibody and anti-β-actin were from Epitomics Burlingame, CA (USA). Anti-Caspase-3 antibody was from Santa Cruz Biotechnology, Inc. Texas (USA). Collagen type-I was from Chrono-log Corporation, Pennsylvania (USA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 1, 1-diphenyl-2-picrylhdrazyl (DPPH) were from HiMedia Laboratories, Mumbai (India). Lactate dehydrogenase (LDH) kit was from AGAPPE diagnostics Ltd., Kerala (India). γ-glutamyl p-nitroanilide and glycylglycine were from Sisco Research laboratories Pvt Ltd., Mumbai (India). All other reagents were of analytical grade.
9
Quantifying Hippocampal p38 MAPK and Caspase-3
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To detect expression of p38 MAPK and cleaved caspase 3 in hippocampal tissues, total protein was first extracted using lysis buffer. Protein extracts were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membranes. The membranes were then blocked and incubated with rabbit monoclonal antiphosphorylated p38 MAPK primary antibody (1:1000 dilution; Santa Cruz, CA, USA) or anti-caspase 3 antibody (1:500 dilution; Santa Cruz, CA, USA), followed by incubation with goat anti-rabbit IgG secondary antibody (1:2000 dilution; Santa Cruz, CA, USA). β-actin was used as a loading control. The expression bands of target proteins were analyzed by Scion Image software (version 4.0.3; Scion Co., Santa Cruz, CA, USA). The densitometric values were used to conduct the statistical analysis. The relative protein expression was calculated over β-actin.
10
Caspase-3 Activity Quantification in C-PAE Cells
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Caspase-3 activity was measured. Briefly, 4 × 105 C-PAE cells were maintained in EMEM medium containing 2% FBS for 24 h. The medium was replaced with fresh medium containing 2% FBS, 10 ng/ml bFGF, and 20 μg/ml ES, ES-BAX, ES-BAK, or ES-BAX-ES. After 24 h, all the cells (adherent or not) were collected and resuspended in SDS-PAGE sample buffer; the proteins were boiled for 5 min, for denaturation. The proteins from the lysates were resolved electrophoretically on a 15% SDS-PAGE and transferred to a nitrocellulose membrane (GE Healthcare, San Diego, CA, USA). The membrane was incubated with anticaspase-3 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 16 h, washed, incubated with a secondary antibody conjugated with HRP for 1 h, and washed again. Immunoreactive bands were visualized using an enhanced chemiluminescence system (Immobilon, Merck Millipore).
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