Cleaved caspase 9

Manufactured by Novus Biologicals

Sourced in China

Cleaved caspase 9 is a laboratory reagent used in the detection and quantification of apoptosis. It is a proteolytic enzyme fragment that is generated during the activation of the intrinsic apoptotic pathway. This fragment can be used as a marker to identify and measure cells undergoing programmed cell death.

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Lab products found in correlation

Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Prolong gold antifade solution, by Thermo Fisher Scientific (1 mentions) Cleaved caspase 3, by Abcam (1 mentions) Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Bcl2 associated x (bax), by Abcam (1 mentions) Apaf 1, by Abcam (1 mentions) X ray image film processor, by Kodak (1 mentions) Goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Ecl system, by CWBIO (1 mentions) Cleaved parp, by Novus Biologicals (1 mentions) Tom20, by Abcam (1 mentions) Affinipure goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)

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Caspase 9 (5 citations)

3 protocols using cleaved caspase 9

Immunohistochemical and Immunofluorescent Analysis of Apoptosis and Eosinophils in Esophageal Tissue

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Serial paraffin sections from tissues fixed in Fekete’s acid alcohol formalin were processed using a Ventanaautostainer (Tucson, AZ). Cleaved caspase 3 (Cell Signalling Technologies; Danvers, MA) and cleaved caspase 9 (Novus Biologicals; Littleton, CO) were used to evaluate apoptosis in the esophagus, and anti-major basic protein (MBP) antibody (obtained from J. Lee, Mayo Clinic, Scottsdale, AZ) to identify eosinophils. Diaminobenzidine (Sigma, St. Louis, MO) was used as chromogen.
For immunofluorescent labeling, frozen sections of esophagus were fixed in 100% acetone and incubated with fluorochrome-labeled antibodies against CD11B (Biolegend) or rabbit antibodies against CHIL3/4 proteins (Stemcell Technologies, Vancouver, CA) followed by Alexafluor 488-labeled donkey anti-rabbit IgG (Jackson Immunoresearch, Westgrove, PA). The sections were cover slipped with ProLong^® Gold Antifade solution (Thermo Fisher Scientific, Grand Island, NY) with 4′,6-diamidino-2-phenylindole (DAPI).

Chien S.J., Silva K.A., Kennedy V.E., HogenEsch H, & Sundberg J.P. (2015). The pathogenesis of chronic eosinophilic esophagitis in SHARPIN-deficient mice. Experimental and molecular pathology, 99(3), 460-467.

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Apoptosis Signaling Pathway Analysis

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Protein lysates were separated by SDS-PAGE in a 10% polyacrimide gel, then transferred to a polyvinylidene difluoride membrane (Millipore, USA). Membranes were incubated with antibody against human Bcl-w (Abcam; 1:200), Apaf-1 (Abcam; 1:500), cleaved Caspase-9 (Novus Bio; 1:1,000), cleaved Caspase-3 (CST; 1:1,000), Bax (Abcam; 1:2,000); or GAPDH (KangChen Bio-tech Inc., China). Peroxidase-conjugated goat anti-mouse IgG (H+L) (Southern Biotech) was used as the secondary antibody. The intensity of staining was visualized using an X-ray image film processor (Kodak, Japan).

Liu S.H., Wang P.P., Chen C.T., Li D., Liu Q.Y., Lv L., Liu X., Wang L.N., Li B.X., Weng C.Y., Fang X.S., Cao X.F., Mao H.B., Chen X.J., Luo S.L., Zheng S.X., Liu G.L, & Wu Y. (2020). MicroRNA-148b enhances the radiosensitivity of B-cell lymphoma cells by targeting Bcl-w to promote apoptosis. International Journal of Biological Sciences, 16(6), 935-946.

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Characterization of Mitochondrial Proteins

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Western blot analysis was performed as detailed previously (40 (link),45 (link),46 (link)). Twenty micrograms of total cell proteins obtained from various cell lines were denatured and loaded on sodium dodecyl sulfate polyacrylamide gels. The gels were electroblotted onto a polyvinylidene difluoride membrane for hybridization. The antibodies obtained from different companies were as follows: Abcam [TOM20 (ab56783), ND1 (ab74257), ND5 (ab92624), CO1 (ab695), CO2 (ab110258), ND6 (ab81212), Total OXPHOS Human WB Antibody Cocktail (ab110411) and CYTC (ab13575)], Cell Signaling Technology [Cleaved Caspase 3 (#9664), Cleaved Caspase 9 (#7237), Cleaved PARP (#5625), SOD1 (4266T), SOD2 (13141T) and Catalase (12980T)], Novus [ND4 (NBP2-47365)] and Proteintech [CYTB (55090-1-AP), ATP8 (26723-1-AP), CO3 (55082-1-AP), Afg3l2 (14631-1-AP), Clpp (15698-1-AP), Lonp1 (66043-1-AP) and GAPDH (10494-1-AP)]. Peroxidase AffiniPure goat anti-mouse IgG and goat anti-rabbit IgG (Jackson) were used as a secondary antibody and protein signals were detected using the ECL system (CWBIO). Quantification of density in each band was performed as detailed previously (40 (link),45 (link),46 (link)).

Xiao Y., Wang M., He Q., Xu L., Zhang Q., Meng F., Jia Z., Zhang F., Wang H, & Guan M.X. (2020). Asymmetrical effects of deafness-associated mitochondrial DNA 7516delA mutation on the processing of RNAs in the H-strand and L-strand polycistronic transcripts. Nucleic Acids Research, 48(19), 11113-11129.

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