Casp7

Manufactured by Cell Signaling Technology

Sourced in United States

CASP7 is a laboratory product offered by Cell Signaling Technology. It is a recombinant protein that represents the active form of caspase-7, a cysteine-aspartic acid protease involved in the execution phase of cell apoptosis. The product is intended for use in research applications.

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Cleaved Casp7 (3 citations)

4 protocols using casp7

Western Blot Antibody Purchasing

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Antibodies against CASP2, CASP3, CASP7, PARP, EZH2 and P53 were purchased from Cell Signaling Technology (Danvers, BSN, USA). Antibody against GAPDH was purchased from Santa Cruz (Dallas, TX, USA). Antibody against BCL-2 was purchased from Proteintech (Chicago, IL, USA). Antibody against LSD1 was purchased from Abcam (Cambridge, UK).

Tang L., Shen H., Li X., Li Z., Liu Z., Xu J., Ma S., Zhao X., Bai X., Li M., Wang Q, & Ji J. (2016). MiR-125a-5p decreases after long non-coding RNA HOTAIR knockdown to promote cancer cell apoptosis by releasing caspase 2. Cell Death & Disease, 7(3), e2137-.

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Western Blot Analysis of Cellular Markers

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Western blotting was conducted as previously described 34 (link). The following primary antibodies were used in this study: GRP78 (# 3177; Cell Signaling Technology), GRP94 (#3196-1; Epitomics), pERK1/2 (#sc-377400, Santa Cruz Biotechnology), p65 (#sc-8008; Santa Cruz Biotechnology), CHOP(#2895; Cell Signaling Technology), cleaved PARP (#sc-56196, Santa Cruz Biotechnology), NLRP3 (#sc-134306; Santa Cruz Biotechnology), HXK2 (#sc-374091, Santa Cruz Biotechnology), IL-1β (#sc-12742; Santa Cruz Biotechnology), IL-18 (#sc133127; Santa Cruz Biotechnology), p-eIF2a (#33981; Cell Signaling Technology), Casp1 (#YH050707C; Epitonics), ZDHHC1 (#AP10315a; Abgent), E-cadherin (#sc8426; Santa Cruz Biotechnology), Occludin (#TA306787; Origene), N-cadherin (#610920; BD transduction laboratories), Vimentin (#5741s; Cell Signaling Technology), G6PD (#12263s; Cell Signaling Technology), Casp3 (#9664; Cell Signaling Technology), Casp7 (#8438; Cell Signaling Technology), GLUT1(#sc-377228; Santa Cruz Biotechnology), beta-actin (#sc8432; Santa Cruz Biotechnology), GAPDH (#sc-47727; Santa Cruz Biotechnology).

Le X., Mu J., Peng W., Tang J., Xiang Q., Tian S., Feng Y., He S., Qiu Z., Ren G., Huang A., Lin Y., Tao Q, & Xiang T. (2020). DNA methylation downregulated ZDHHC1 suppresses tumor growth by altering cellular metabolism and inducing oxidative/ER stress-mediated apoptosis and pyroptosis. Theranostics, 10(21), 9495-9511.

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Quantification of Apoptosis Markers in CAD

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Western blots were performed in according to standard procedures. HUVECs were seeded in 6-well plates and lysed with 100 μl of a cell lysis solution containing 2 μl of phenylmethanesulfonyl fluoride buffer. Equal amounts of protein (40 μg) were loaded onto 12% SDS–PAGE gels (Millipore). Primary antibodies for Casp7 (1:1000 dilution; Cell Signaling), Casp3 (1:1000 dilution; Cell Signaling), Apaf1 (1:1000 dilution; Cell Signaling) and PTEN (1:1000 dilution; Cell Signaling) were used. The antibody for β-actin (1:1000 dilution; Cell Signaling) was used as the endogenous control. The gels were run under the same experimental conditions and the original whole gel blots were included in the supplementary data. The levels of TNF-α, Apaf-1 and Casp7 protein in the serum samples obtained from the patients with CAD were measured by Enzyme-Linked Immunosorbent Assay Kits (Elabscience).

Tang Y., Zhang Y.C., Chen Y., Xiang Y., Shen C.X, & Li Y.G. (2015). The role of miR-19b in the inhibition of endothelial cell apoptosis and its relationship with coronary artery disease. Scientific Reports, 5, 15132.

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Western Blot Analysis of Cellular Signaling

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Untreated as well as transfected whole cell lysates were prepared using the cOmplete Lysis-M kit (Roche Applied Science, Indianapolis, IN). Protein samples were prepared by combining lysates with NuPAGE Sample Reducing Agent, NuPAGE LDS Sample Buffer, and nuclease-free water (Life Technologies) and then boiling for 5 minutes at 100°C. Samples were loaded onto NuPAGE 4-12% Bis Tris Gels (Life Technologies), separated by SDS-PAGE, and then transferred to PVDF membranes. Proteins were blocked in 5% non-fat milk and incubated with the appropriate antibody in 5% BSA (Sigma-Aldrich). The following antibodies were used for Western blot analyses: AKT (#9272), pAKT Thr308 (#4056), pAKT Ser473 (#4058), CASP7 (#9492), Fox01 (#2880), pFox01 Ser256 (#9461), GSK-3β (#9315), pGSK-3β Ser9 (#9336), INNPL1 (#2839), p70 S6 Kinase (#2708), PDK1 (#5662), pPDK1 Ser241 (#3438), PI3K p85 (#4292), and GAPDH (#2118) [Cell Signaling, Danvers MA]; and pGSK-3β Tyr216 (#75745) [Abcam Inc., Cambridge, MA]. Protein bands were visualized using Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, Rockford, IL).

Shanmugam V., Ramanathan R.K., Lavender N.A., Sinari S., Chadha M., Liang W.S., Kurdoglu A., Izatt T., Christoforides A., Benson H., Phillips L., Baker A., Murray C., Hostetter G., Von Hoff D.D., Craig D.W, & Carpten J.D. (2014). Whole genome sequencing reveals potential targets for therapy in patients with refractory KRAS mutated metastatic colorectal cancer. BMC Medical Genomics, 7, 36.

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