A rabbit monoclonal anti-FLAG (1:50, Cell Signaling Technology, Danvers, MA) was used in co-immunoprecipitation experiments, whereas Western blots were probed with rabbit polyclonal anti-LC3 (Cell Signaling Technology), rabbit polyclonal anti-β actin (Cell Signaling Technology), rabbit polyclonal anti-p62 (Sigma), or mouse monoclonal anti-myc (Thermo Scientific, Waltham, MA) (all 1:1,000). Densitometry was undertaken with Image J software (Schneider et al., 2012 (link)).
Rabbit polyclonal anti lc3
Manufactured by Cell Signaling Technology
Sourced in United Kingdom, United States
Rabbit polyclonal anti-LC3 is an antibody product developed by Cell Signaling Technology. The antibody targets the LC3 protein, which is a key component of the autophagy process in cells.
Automatically generated - may contain errors
Lab products found in correlation
Polyclonal rabbit anti β actin, by Cell Signaling Technology (1 mentions)
Mouse monoclonal anti myc, by Thermo Fisher Scientific (1 mentions)
Anti flag, by Cell Signaling Technology (1 mentions)
Mouse monoclonal anti β actin, by Merck Group (1 mentions)
Ecl prime, by GE Healthcare (1 mentions)
Rapamycin, by Merck Group (1 mentions)
Cyto id autophagy detection kit, by Enzo Life Sciences (1 mentions)
3 methyladenine 3 ma, by Merck Group (1 mentions)
3 protocols using rabbit polyclonal anti lc3
1
Immunoprecipitation and Western Blot Analysis
2
Immunoblotting for Protein Analysis
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Biochemical analysis was performed as previously described [13 (link),15 (link)]. For immunoblotting, rabbit monoclonal anti-alpha-synuclein (1:5000) was purchased from Abcam (Cambridge Biomedical Campus, Cambridge, UK), rabbit polyclonal anti-LC-3 (1:1000) was purchased from Cell Signaling Technology (Danvers, MA, USA), and mouse monoclonal anti-β-actin (1:5000) was purchased from Sigma-Aldrich (St. Louis, MO, USA). After the primary antibody reaction, the membrane was then incubated with the following secondary antibodies at RT for 30 min: goat anti-rabbit IgG antibody, peroxidase conjugated, H+L (Merck KGaA, Darmstadt, Germany) (1:2000 dilution), or goat anti-mouse IgG antibody, peroxidase conjugated, H+L (Merck KGaA) (1:2000 dilution). Finally, the membrane was incubated with ECL prime (GE Healthcare) to generate chemiluminescence. Chemiluminescence was detected by using a Fusion System (Vilber-Lourmat, Marne-la-Vallée, France). The band density was measured by ImageJ software (NIH, New York, NY, USA).
3
Autophagy Regulation in Cancer Cells
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3-Methyladenine (3-MA) and rapamycin were purchased from Sigma; Merck KGaA (Darmstadt, Germany). A Cyto-ID autophagy detection kit was purchased from Enzo Life Sciences, Inc. (Farmingdale, NY, USA; cat. no. ENZ-51031-K200). HCPT was purchased from Bailingwei Technology Co., Ltd. (Beijing, China) and an MTT cell viability assay kit was purchased from Zhejiang Tianshun Biotechnology Co., Ltd. (Zhejiang, China; http://tianshunbiotech.com/index_en.asp ). An Annexin V-propidium iodide (PI) apoptosis kit was purchased from Yeasen Biotechnology Co., Ltd. (Shanghai, China). Rabbit polyclonal anti-Beclin-1 (cat. no. 4122), rabbit polyclonal anti-phosphorylated mammalian target of rapamycin (p-mTOR) (cat. no. 5536), rabbit polyclonal anti-Bcl-2-associated X protein (Bax) (cat. no. 2772), rabbit polyclonal anti-Bcl-2 (cat. no. 2876), rabbit polyclonal anti-GAPDH (cat. no. 5174), goat anti-rabbit immunoglobulin secondary antibody (cat. no. 14708), Tubulin antibody (cat. no. 2146) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA); Rabbit polyclonal anti-LC3 (cat. no. L7543) were purchased from Sigma; Merck KGaA (Darmstadt, Germany).
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