Apaf 1

Manufactured by Proteintech

Sourced in Canada, United States

Apaf-1 is a protein involved in the regulation of apoptosis, or programmed cell death. It plays a central role in the initiation of the intrinsic apoptotic pathway. Apaf-1 forms a multi-protein complex called the apoptosome, which activates caspase enzymes and leads to the execution of cell death.

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Lab products found in correlation

Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Lamc1, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated anti rabbit na 934v, by GE Healthcare (1 mentions) Anti mouse na931v, by GE Healthcare (1 mentions) Protease inhibitor, by Merck Group (1 mentions) Caspase 9, by Proteintech (1 mentions) Srebp1, by Beyotime (1 mentions) P erk, by Bioss Antibodies (1 mentions) P ire1, by Bioss Antibodies (1 mentions) Caspase 3, by Proteintech (1 mentions) Gapdh, by Proteintech (1 mentions) Bcl2 associated x (bax), by Proteintech (1 mentions) Bcl 2, by Proteintech (1 mentions) Cleaved casp3, by Affinity Biosciences (1 mentions) Bcl2 associated x (bax), by Affinity Biosciences (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Gapdh, by ZSGB-BIO (1 mentions) Bcl 2, by Affinity Biosciences (1 mentions)

3 protocols using apaf 1

Western Blot Analysis of Protein Targets

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Cells were homogenised in lysis buffer [10-mM Tris-HCl (pH 7.4), 150-mM NaCl, 1% NP-40, 10-mM NaF, 0.2-mM Na₃VO₄ and protease inhibitor (Sigma–Aldrich)]. Proteins (30 μg) were fractionated on 10% SDS-PAGE, blotted onto a polyvinylidene difluoride membrane (Millipore, MA) and probed with primary antibodies, including SRF, LAMC1 (Santa Cruz, CA) and Apaf-1 (Proteintech, Wuhan). GAPDH (ZSGQ-BIO, Beijing) was used as a loading control. The secondary antibodies were HRP-conjugated anti-rabbit (NA 934V) and anti-mouse (NA 931V) antibodies (GE Healthcare Life Sciences, MA). Proteins were visualised by chemiluminescence according to the manufacturer's instructions (Thermo, PA), followed by exposure to X-ray film. The density of bands was densito-metrically quantified using ImageJ (National Institutes of Health, MD).

Xu D., Guo Y.B., Zhang M, & Sun Y.Q. (2018). The subsequent biological effects of simulated microgravity on endothelial cell growth in HUVECs. Chinese Journal of Traumatology, 21(4), 229-237.

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Comprehensive Western Blot Analysis of Metabolic and Stress Signaling

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The protein extraction and western blot were performed according to the methods previously described [24 (link)]. The primary antibodies include SREBP1 (1:1000, Beyotime), ACC (1:1000, Proteintech), p-ACC (Ser⁷⁹, 1:1000, Proteintech), AMPK (1:1000, Beyotime), p-AMPK (Ser¹⁷², 1:1000, CST), Protein kinase RNA-like endoplasmic reticulum kinase (PERK, 1:1000, Bioss), p-PERK (Thr⁹⁸⁰, 1:1000, Bioss), Phosphorylated eukaryotic initiation factor 2 (p-EIF2α at Ser⁵¹, 1:1000, Beyotime), IRE1 (1:1000, Proteintech), p-IRE1 (Ser⁷²⁴, 1:1000, Bioss), X-box binding protein 1 spliced (XBP1s, 1:1000, Beyotime), Activating transcription factor 4 (ATF4, 1:1000, Proteintech), C/EBP homologous protein (CHOP, 1:1000, Proteintech), p-CHOP (Ser³⁰, 1:1000, Bioss), B-cell lymphoma-2 (Bcl2, 1:1000, Proteintech), Bcl2-associated X (Bax, 1:1000, Proteintech), CytC (1:1000, Proteintech), Apoptotic protease activating factor-1 (Apaf-1, 1:1000, Proteintech), caspase 9 (1:1000, Proteintech), caspase 3 (1:1000, Proteintech), DRP1 (1:1000, Beyotime), p-DRP1 (Ser⁶¹⁶, 1:1000, SAB), MFN1 (1:1000, Proteintech), MFN2 (1:1000, Proteintech), Fission 1 (FIS1, 1:1000, Proteintech), ACTB (1:1000, Beyotime) and GAPDH (1:1000, Proteintech).

Zhong Y., Li Z., Jin R., Yao Y., He S., Lei M., Wang X., Shi C., Gao L, & Peng X. (2022). Diosgenin Ameliorated Type II Diabetes-Associated Nonalcoholic Fatty Liver Disease through Inhibiting De Novo Lipogenesis and Improving Fatty Acid Oxidation and Mitochondrial Function in Rats. Nutrients, 14(23), 4994.

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Western Blot Analysis of Apoptosis Markers

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In brief, an equal amount of protein was separated with 10%–12% SDS-polyacrylamide gel electrophoresis (PAGE) and transferred to 0.22-μm polyvinylidene fluoride (PVDF) membranes (Millipore, Bedford, MA, USA). Thereafter, the membranes were blocked with 5% milk and immunoblotted with primary antibodies. The antibodies included APAF1 (1:500; Proteintech, Chicago, IL, USA), GAPDH (1:2,000; Zsbio, Beijing, China), and apoptotic-associated proteins such as pro- or cleaved-CASP9, cleaved-CASP3, Bcl2, and Bax (1:1,000; Affinity, OH, USA).

Wu H., Hu X., Li Y., Chen Q., Sun T., Qiao Y., Qin W., Wu Z., Fu B., Zhao H., Zhang R, & Wei M. (2020). LNC473 Regulating APAF1 IRES-Dependent Translation via Competitive Sponging miR574 and miR15b: Implications in Colorectal Cancer. Molecular Therapy. Nucleic Acids, 21, 764-779.

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