Cells were lysed in HALT inhibitor-containing RIPA buffer with mild sonication for indicated western blot assays. The mitochondrial fraction was prepared from cell pellets utilizing a reagent-based method per manufacturer’s instruction (Mitochondria Isolation Kit; Pierce), and lysed in RIPA buffer and analyzed with Western blot as previously reported[27 (link)]. Antibodies were from Santa Cruz Biotech (p53/DO-1, Bax/N-20, PCNA/PC10 and p21/C-19), Calbiochem (Mdm2/Ab3), abcam (ubiquitin/ab7780, HSP90/AC88 and cyclin E/ab33911), Dako-Agilent (AR/AR441) and Sigma-Aldrich (Cyclin A/C4710). Anti-β-actin (abcam, ab8227) was included for equivalent protein loading.
Cyclin a c4710
Manufactured by Merck Group
Cyclin A/C4710 is a laboratory equipment product designed for use in various research applications. It serves as a tool for the study of cell cycle regulation. The product's core function is to facilitate the analysis and measurement of cyclin A, a key regulatory protein involved in cell division. This equipment does not include any interpretation or extrapolation of its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Abcam (2 mentions)
Mdm2 ab3, by Abcam (2 mentions)
Ar ar441, by Merck Group (2 mentions)
Bt474, by American Type Culture Collection (1 mentions)
Hcc1419, by American Type Culture Collection (1 mentions)
Las 3000 imaging system, by Fujifilm (1 mentions)
Western lightning, by PerkinElmer (1 mentions)
P21 sc 397, by Santa Cruz Biotechnology (1 mentions)
P27 sc 528, by Santa Cruz Biotechnology (1 mentions)
Cyclin d1 sc 450, by Santa Cruz Biotechnology (1 mentions)
3 protocols using cyclin a c4710
1
Mitochondrial Protein Analysis by Western Blot
2
Mitochondrial Protein Analysis by Western Blot
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Cells were lysed in HALT inhibitor-containing RIPA buffer with mild sonication for indicated western blot assays. The mitochondrial fraction was prepared from cell pellets utilizing a reagent-based method per manufacturer’s instruction (Mitochondria Isolation Kit; Pierce), and lysed in RIPA buffer and analyzed with Western blot as previously reported[27 (link)]. Antibodies were from Santa Cruz Biotech (p53/DO-1, Bax/N-20, PCNA/PC10 and p21/C-19), Calbiochem (Mdm2/Ab3), abcam (ubiquitin/ab7780, HSP90/AC88 and cyclin E/ab33911), Dako-Agilent (AR/AR441) and Sigma-Aldrich (Cyclin A/C4710). Anti-β-actin (abcam, ab8227) was included for equivalent protein loading.
3
Taspase1 Knockdown in Breast Cancer Cell Lines
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BT-474 and HCC1419 cell lines were obtained from American Type Culture Collection and cultivated for no more than 2 months after each frozen aliquot was thawed. Amphotropic retrovirus carrying Taspase1 specific knockdown hairpin was generated as described26 (link),29 (link). To assay cell proliferation, 1 × 105 cells were seeded onto each well of a 6-well plate and counted 4 days later. Cell cycle and cell death analyses were performed as described26 (link). For western blot, cells and tissues were lysed in standard RIPA buffer. The anti-Taspase1 rabbit polyclonal antibody is as described22 (link),26 (link). Antibodies for cyclin E2 (4132, Cell Signaling), cyclin A (C4710, Sigma), p21 (sc-397, Santa Cruz Biotechnology), p27(sc-528, Santa Cruz Biotechnology), cyclin D1(sc-450, Santa Cruz Biotechnology), p16(554079, BD Pharmingen), and ErbB2(OP-15, Calbiochem) were purchased from indicated companies. Antibodies were detected using the enhanced chemiluminescence method (Western Lightning, PerkinElmer). Immunoblot signals were acquired with the LAS-3000 Imaging system (FujiFilm) and were analyzed with ImageJ software.
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