Allograft tumors and the livers and kidneys were dissected and fixed in 10% (v/v) neutral-buffer formalin. They were then dehydrated in ascending grades of ethanol and xylene and embedded in paraffin wax. Thereafter, the sections (4 μm) were cut with a microtome (Leica, Germany). Anti-DNMT1 antibody, anti-caspase-3 antibody and goat anti-rabbit antibody IgG (BOSTER, Wuhan, China) were used for immunostaining. For HE staining, after deparaffinization and rehydration, 4-μm sections were stained with hematoxylin solution for 5 min followed by 5 dips in 1% acid ethanol and then rinsing in water. Next, the sections were stained with eosin solution for 3 min, followed by dehydration with graded alcohol and clearing in xylene.
Anti caspase3 antibody
Manufactured by Boster Bio
Sourced in China
The Anti-caspase3 antibody is a laboratory tool used for the detection and analysis of caspase-3, a key enzyme involved in the process of apoptosis, or programmed cell death. This antibody can be utilized in various immunological techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and localization of caspase-3 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Microtome, by Leica (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hoechst, by Wuhan Servicebio Technology (1 mentions)
S1000 with bestar sybr green rt pcr master mix, by DBI Bioscience (1 mentions)
Anti gapdh antibody, by Wuhan Servicebio Technology (1 mentions)
Anti tgf β1 antibody, by Boster Bio (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Tunel kit, by Roche (1 mentions)
Biotinylated goat anti rabbit igg, by Boster Bio (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Ix51 microscope, by Olympus (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Protein lysis buffer, by Beyotime (1 mentions)
4 protocols using anti caspase3 antibody
1
Immunohistochemical Analysis of Allograft Tumors
2
Investigating FNDC3B in Kidney Cell Apoptosis
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C57 mice (weight 20–25 g, male) were obtained from the Changzhou CAVENS Laboratory Animal (Jiangsu, China). HK-2 cells were acquired from the BeNa Culture Collection (Beijing, China). The following materials were also used: anti-FNDC3B antibody (Atlas Antibodies AB, Sweden); anti-TGF-β1 antibody (Boster, Wuhan, China); anti-GAPDH antibody (Service Bio, Wuhan, China); anti-caspase3 antibody (Boster, Wuhan, China); TRIzol (Invitrogen, USA); fetal bovine serum (Gibco, USA); TUNEL Kit (Roche, Shanghai, China); Hoechst (Service Bio, Wuhan, China); Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA); Bio-Rad S1000 with Bestar SYBR Green RT-PCR Master Mix (DBI Bioscience, Shanghai, China); and AAV GPAAV-CMV-MCS-EF1-ZsGreen1-WPRE (Genomeditech Co., LTD, Shanghai, China).
3
Immunohistochemical Analysis of Caspase-3 Expression
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After deparaffinization and rehydration, immunohistochemical staining of paraffin-embedded tumor tissue sections was performed using UltraSensitiveTM S-P IHC Kit according to the manufacturer’s protocols. Briefly, the sections were incubated with anti-Caspase 3 antibody (1:100; Boster Biological Technology, Ltd., Wuhan, China) at 4°C overnight and reincubated with biotinylated goat-anti-rabbit IgG (1:100; Boster Biological Technology, Ltd.) for 30 minutes. After washing in PBS three times for 5 minutes, the signal was visualized using 0.05% diaminobenzidine substrate and counterstained with hematoxylin. Thereafter, the sections were examined and microphotographed under an Olympus IX51 microscope (400×; Olympus Corporation). Brown cytoplasmic staining was considered to be positive for Caspase 3, and the relative optical density of Caspase 3 was calculated as integral optical density (IOD)treated group/IODcontrol group by using Image-pro plus 6.0 (Media Cybernetics, Silver Spring, MD, USA).
4
Western Blot Analysis of Caspase-3
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After incubating with control, 10 and 100 µg/ml nPANI for 24 hours, cells were lysed with protein lysis buffer (Beyotime, China). The concentration of total protein was detected by Lowry method. Equal amounts of protein were fractionated by 10% SDS gels and transferred to polyvinylidene difluoride membranes (Millipore Corporation, USA). After blocked with 5% nonfat dry milk, the membranes were incubated with an anti-caspase3 antibody (1∶1000 dilution, Boster, China) or anti-actin antibody (1∶1000 dilution, Boster, China) at 4°C overnight. After removing primary antibodies, the membranes were washed 3 times for 5 minutes by TBST (Tris-Buffered Saline, 0.1% Tween-20) solution and followed by exposure of secondary antibodies (1∶5000 dilution, goat anti-rabbit or goat anti-mouse, Boster, China) for 2 hours at 4°C. Finally, after wash, bands on the membranes were visualized by developer and fixing solution [21] (link). The protein bands were quantified using the ImageJ software (NIH).
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