Rabbit anti cleaved casp3

Manufactured by Cell Signaling Technology

Rabbit anti-cleaved Casp3 is a primary antibody that recognizes the cleaved form of Caspase-3 enzyme. Caspase-3 is a key effector of apoptosis, or programmed cell death. This antibody can be used to detect and study the activation of Caspase-3 in various biological samples.

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Lab products found in correlation

4oh tmx, by Merck Group (1 mentions) Rapamycin, by LC Laboratories (1 mentions) Dapi fluoromount g, by Southern Biotech (1 mentions) Af647 donkey anti rabbit, by Jackson ImmunoResearch (1 mentions) Protease inhibitor and phosphatase inhibitor cocktail, by Roche (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Sodium dodecyl sulfate (sds), by Merck Group (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Sodium deoxycholate, by Merck Group (1 mentions) Np 40, by Merck Group (1 mentions) Tx 160, by TOMY (1 mentions) Mouse anti β actin, by Merck Group (1 mentions)

Spelling variants of this product

Casp3 (38 citations) Cleaved CASP3 (24 citations) Anti-CASP3 (14 citations) Anti-cleaved Casp3 (11 citations) Rabbit anti-CASP3 (6 citations)

3 protocols using rabbit anti cleaved casp3

Tracheosphere Formation and Regulation

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Epithelial cells from wild-type mice were used to induce tracheospheres formation. After 10 days, Rapamycin (final concentration 200nM, LC Laboratories) or EtOH 100% (control) was added to the MTEC/SF for 6 days. Medium was changed every day.
For TSC1 KO recombination in culture, epithelial cells from Krt5-Cre Rosa26R-LacZ with or without TSC1 fl/fl were used to induce spheres formation. After 9 days, 4-OH-Tmx (final concentration 300nM, Sigma) or EtOH 100% (control) was added to the MTEC/SF for 48h.
Then tracheospheres were fixed with 4% paraformaldehyde in PBS, stained and imaged as described above for whole trachea. Antibody used to detect cleaved Caspase-3 was rabbit anti-cleaved Casp3, 1:400 (#9661, Cell Signaling).

Haller S., Kapuria S., Riley R.R., O’Leary M.N., Schreiber K.H., Andersen J.K., Melov S., Que J., Rando T.A., Rock J., Kennedy B.K., Rodgers J.T, & Jasper H. (2017). mTORC1 activation during repeated regeneration impairs somatic stem cell maintenance. Cell stem cell, 21(6), 806-818.e5.

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Immunofluorescent Analysis of Mouse Tumors

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Mouse tumors were embedded for 4 hours in 4% paraformaldehyde overnight in 30% sucrose and mounted in OCT medium. Fifty-micrometer-thick sections were cut and fixed in 4% paraformaldehyde for 20 min. After washing and permeabilization [0.5% PBS/BSA + Triton X-100 (1/400e, AppliChem); 20 min, room temperature], sections were stained overnight at 4°C using a rat anti–Lyve-1 antibody (1/300e; Novus) and a rabbit anti–cleaved casp-3 (1/100e; Cell Signaling Technologies). Alexa Fluor 546–labeled goat anti-rat (1/1000e; Invitrogen) and donkey anti-rabbit AF647 (1/1000e; Jackson ImmunoResearch) and Alexa Fluor 488–labeled anti-CD45.1 (1/100e; BioLegend, clone A20) antibodies were incubated for 2 hours at room temperature. Sections were mounted with DAPI Fluoromount-G (SouthernBiotech).

Garnier L., Pick R., Montorfani J., Sun M., Brighouse D., Liaudet N., Kammertoens T., Blankenstein T., Page N., Bernier-Latamani J., Tran N.L., Petrova T.V., Merkler D., Scheiermann C, & Hugues S. (2022). IFN-γ–dependent tumor-antigen cross-presentation by lymphatic endothelial cells promotes their killing by T cells and inhibits metastasis. Science Advances, 8(23), eabl5162.

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Immunoblotting of Cleaved Caspases

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IB was performed as described before [32 (link)]. Briefly, top-most portion of left lungs were excised and placed in ice-cold HBSS. Equal weight of tissue (approximately 100 mg of tissue from each mouse) were sonicated in RIPA (25 mM Tris, 150 mM sodium chloride [Sigma], 1% NP-40 [Sigma], 1% sodium deoxycholate [Sigma], and 0.1% SDS [Sigma], pH 7.6; 200 μl per sample) supplemented with protease inhibitor and phosphatase inhibitor cocktail (Roche). After sonication, tissues were lysed on ice for 30 mins, collected by centrifugation at 150,000 rpm at 4°C for 20 mins using a Tomy TX-160 high speed refrigerated micro centrifuge. The resulting supernatant (tissue lysate) was collected for IB analysis. Beta actin levels were used to determine comparable protein quantities in all samples for IB. Proteins were transferred to PVDF membranes (Bio-Rad), treated with chemiluminescence reagent (Clarity, Bio-Rad) for signal development and imaged on Kwik Quant Gel Imaging System (Kindle Biosciences Inc). Antibodies used were rabbit anti-cleaved Casp8 (8592, Cell Signaling Technology), rabbit anti-cleaved Casp3 (9661, Cell Signaling Technology) and mouse anti-β-actin (A2228, Sigma).

Mandal P., Lyons J.D., Burd E.M., Koval M., Mocarski E.S, & Coopersmith C.M. (2021). Integrated evaluation of lung disease in single animals. PLoS ONE, 16(7), e0246270.

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