The following primary antibodies were used for immunohistochemistry: rabbit anti-active caspase-3 (1:500; Abcam, Cambridge, United Kingdom), rabbit anti-DCX (1:500; Cell Signaling Technology, Danvers, MA), rabbit anti-glial fibrillary acidic protein (GFAP) (1:500; Agilent, Santa Clara, CA), goat anti-Olig2 (1:50; R&D Systems, Minneapolis, MN), rabbit anti-Iba1 (1:500; Fuji-Wako, Osaka, Japan), rabbit anti-Ki67 (1:500; Abcam), goat anti-MBP (1:500; Santa Cruz Biotechnology, Dallas, TX), rat anti-MBP (1:1,000; Abcam), mouse anti-NeuN (1:100; Merck-Millipore, Burlington, MA), and rat anti-NG2 (1:500; Abcam). The conjugated secondary antibodies were donkey anti-rabbit, anti-rat, anti-mouse, and anti-goat antibodies (Alexa Fluor 488, 568, and 647; Thermo Fisher Scientific, Waltham, MA) at a dilution of 1:500. For the EdU reaction, azide-modified dyes (Alexa Fluor 488 and 594 azide; Thermo Fisher Scientific) were used.
Rabbit anti active caspase 3
Manufactured by Abcam
Rabbit anti-active caspase-3 is a primary antibody that binds to the active form of caspase-3, a key enzyme involved in the execution phase of apoptosis. This antibody can be used to detect and quantify the level of active caspase-3 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti glial fibrillary acidic protein, by Agilent Technologies (1 mentions)
Goat anti olig2, by R&D Systems (1 mentions)
Rabbit anti dcx, by Cell Signaling Technology (1 mentions)
Goat anti mbp, by Santa Cruz Biotechnology (1 mentions)
Mouse anti neun, by Merck Group (1 mentions)
Rabbit anti ki67, by Abcam (1 mentions)
Rat anti mbp, by Abcam (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions)
Lsm 710, by Zeiss (1 mentions)
Mouse anti brdu, by BD (1 mentions)
Mouse anti repo, by Developmental Studies Hybridoma Bank (1 mentions)
Chicken anti tuj1, by Merck Group (1 mentions)
Rabbit anti ng2, by Merck Group (1 mentions)
Mouse anti olig2, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Chicken anti vimentin, by Merck Group (1 mentions)
80i fluorescence microscope, by Nikon (1 mentions)
Mouse anti gapdh, by Cell Signaling Technology (1 mentions)
Rabbit anti lc3, by Abcam (1 mentions)
6 protocols using rabbit anti active caspase 3
1
Immunohistochemistry for Neural Cell Markers
2
Caspase-3 Activation in hBMVEC
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hBMVEC were exposed for 6 h to cytokines and then fixed. After 0.2% Triton-X permeabilization they were incubated with primary antibody, rabbit anti active caspase-3 (1:100, Abcam) for 5 h at room temperature. F-actin staining (Alexa Fluor 488 Phalloidin, Life Technologies), performed on fixed cells, was executed according to the manufacturer’s procedure.
3
Immunofluorescence Staining of Drosophila Larval Brains
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The following antibodies were used: Guinea pig anti-β-galactosidase (1:500, a gift from Y. Hiromi), rat anti-β-galactosidase (1:500, a gift from Y. Hiromi), rabbit anti-β-galactosidase (1:500, Cappel), mouse anti-Repo (1:200, Developmental Studies Hybridoma Bank (DSHB) 8D12), mouse anti-Chaoptin (1:100, DSHB, 22B10), rat anti-Miranda (Mira) (1:100), guinea pig anti-Deadpan (Dpn) (1:1000), mouse anti-BrdU (1:250, Becton, Dickinson and Company), mouse anti-Dlp (1:4, DSHB, 13G8), rabbit anti-active caspase 3 (1:500, Abcam), mouse anti-Svp (1:4, a gift from Y. Hiromi), rabbit anti-pH3(1:1000, Upstate Biotechnology), and Alexa Fluor 488, 546, 633 conjugated secondary antibodies (1:500, Invitorgen).
Larval brains were dissected in PBS and fixed with 3.7% paraformaldehyde in PBS for 30 min at room temperature. After washing with PBST (0.1% triton X-100 in PBS), tissues were blocked with 5% normal goat serum in PBST for 30 min. The samples were incubated with primary antibodies at 4 °C overnight. After washing with PBS, the samples were incubated with secondary antibodies for 2 hours at room temperature. The tissues were mounted in Vectashield (VectaLabs) and fluorescent signals were observed by confocal laser scanning microscope (Zeiss LSM710) with a 40X water immersion objective.
Larval brains were dissected in PBS and fixed with 3.7% paraformaldehyde in PBS for 30 min at room temperature. After washing with PBST (0.1% triton X-100 in PBS), tissues were blocked with 5% normal goat serum in PBST for 30 min. The samples were incubated with primary antibodies at 4 °C overnight. After washing with PBS, the samples were incubated with secondary antibodies for 2 hours at room temperature. The tissues were mounted in Vectashield (VectaLabs) and fluorescent signals were observed by confocal laser scanning microscope (Zeiss LSM710) with a 40X water immersion objective.
4
Immunocytochemical Analysis of Neural Cells
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Cells from neurosphere differentiation cultures were fixed in 4% PFA with a 30% sucrose solution for 30 min at 37°C. For immunocytochemistry, cultures were preincubated for 1 h in blocking solution (10% goat serum, 0.1% Triton X-100, BSA), followed by overnight incubation with the appropriate primary antibody at 4°C. The following primary antibodies were used: mouse anti-hGFAP (1:500, Sternberger Monoclonal), chicken anti-vimentin (1:200, Millipore), rabbit anti-NG2 (1:200, Millipore), mouse anti-Olig2 (1:200, Millipore), chicken anti-Tuj1 (1:200, Millipore), rabbit anti-active caspase-3 (1:200, Abcam), rabbit anti-HSP27 (1:200, Abcam), and rabbit anti-cathepsin (1:200, Abcam). The corresponding secondary antibodies were incubated for 2 h (Alexa-Fluor 405, 488, 555, or 647 goat anti-mouse, chicken or rabbit; 1:500; Invitrogen), followed by incubation with DAPI (1:1,000, Sigma) for 10 min and rinsing before being mounted on glass slides with Fluorsave (Calbiochem). Analyses were performed with a Nikon 80i fluorescence microscope at 40× or 63× magnification.
5
Western Blot Analysis of Apoptosis Markers
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The cells were lysed with a lysis buffer (Beyotime, Haimen, China), and centrifuged at 12,000 × g for 20 min. The total protein concentration was determined using the a BCA protein assay kit (Pierce). Samples were resolved in SDS-PAGE, transferred to 0.45 μm nitrocellulose transfer membranes and analyzed separately. After blocking with 5 % skim milk at room temperature for 60 min, the blots were survey with primary antibodies against mouse anti-Bcl-2 (1:500; Abcam), rabbit anti-LC3 (1:1000; Abcam), mouse anti-GAPDH (1:1000; Cell Signal), and rabbit anti-active caspase-3 (1:1000; Abcam), at 4 °C for overnight. We washed the membranes three times with TBST buffer (20 mmol/L Tris-buffered saline and 0.1 % Tween 20) for 1 h. And we used peroxidase conjugated anti-mouse-IgG/anti rabbit-IgG as secondary antibodies. We did Chemiluminescence with Amersham ECL plus Western blotting detection system (GE healthcare). After washing with the TBST buffer, the membranes were measured with the Odyssey Infrared Imaging System (LI-COR).
6
Immunohistochemical Analysis of Zebrafish Fin Regeneration
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Fins were fixed in 4% paraformaldehyde in PBS, embedded in OCT, and cryosectioned. Antibody staining was performed as previously described [22 (link)]. The following primary antibodies were used: rat anti-BrdU (1:200), rabbit anti-active-caspase 3 (1:10000) (both Abcam), rabbit anti-Tenascin C (1:500; US Biological), mouse anti-Zns5 (1:100; Zebrafish International Resource Center), rabbit anti-And1 (1:5000; Eurogentec). The following secondary antibodies were used at a concentration of 1:500: goat anti-rat Alexa Fluor 488 (Molecular Probes), and goat anti-rabbit Cy3-conjugated and anti-mouse Cy5-conjugated antibodies (Jackson ImmunoResearch).
For proliferation assay, BrdU-positive cells distal to the amputation plane were counted in the mesenchyme and epidermis, and the number of BrdU-positive cells was normalized to the total number of DAPI-stained nuclei. Fluorescent pictures were taken with a confocal microscope (TCS SP5; Leica) and Image J 1.43q software was used for the measurements.
For proliferation assay, BrdU-positive cells distal to the amputation plane were counted in the mesenchyme and epidermis, and the number of BrdU-positive cells was normalized to the total number of DAPI-stained nuclei. Fluorescent pictures were taken with a confocal microscope (TCS SP5; Leica) and Image J 1.43q software was used for the measurements.
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