Cells were seeded and differentiated in Costar 25 cm2 flasks (Corning) at 1.2 million cells/well. After incubation, cells were scrapped in 150 μL lysis buffer (40 mM Tris pH 7.5, 50 mM potassium chloride, 1 mM EDTA, 1% sodium dodecyl sulfate) containing a protease inhibitor mixture (1 tablet in 2 mL water added at a 1/25 dilution; Hoffman-La Roche Ltd, Basel, Switzerland) and phosphatase inhibitors (sodium metavanadate 25 mM, 4-nitrophenyl phosphate 250 mM, β-glycerophosphate 250 mM, sodium fluoride 125 mM at a 1/25 dilution). Western blot analysis was performed as described in Notte et al24 (link) using a rabbit monoclonal anti-phosphorylated eIF2α antibody (119A11; Cell Signaling Technology, Inc., Danvers, MA, USA), a mouse anti-β-actin antibody (A5441; Sigma), a rabbit polyclonal anti-cleaved caspase 3 antibody (9662S; Cell Signaling), a rabbit polyclonal anti-caspase 7 antibody (9492; Cell Signaling), a rabbit polyclonal anti-caspase 4 antibody (4450S; Cell Signaling), a rabbit polyclonal anti-caspase 9 antibody (9502; Cell Signaling), a mouse monoclonal anti-caspase 8 antibody (9746; Cell Signaling), a mouse monoclonal anti-LC3 antibody (Nanotools GmbH, Munchen, Germany), or a mouse anti-p62 antibody (Abnova, Taipei, Taiwan).
Mouse anti p62 antibody
Manufactured by Abnova
Sourced in Germany
The Mouse anti-p62 antibody is a primary antibody that recognizes the p62 protein, also known as sequestosome-1 (SQSTM1). The p62 protein is involved in the regulation of various cellular processes, including autophagy, inflammation, and cell signaling. This antibody can be used for the detection and analysis of the p62 protein in biological samples using techniques such as Western blotting, immunohistochemistry, and immunocytochemistry.
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Lab products found in correlation
Mouse anti β actin antibody, by Merck Group (1 mentions)
Protease inhibitor mixture, by Roche (1 mentions)
Rabbit anti lc3 antibody, by Novus Biologicals (1 mentions)
Mouse anti gapdh antibody, by Merck Group (1 mentions)
Anti flag m2 hrp antibody, by Merck Group (1 mentions)
Anti beclin 1 antibody, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti tom20 antibody, by Santa Cruz Biotechnology (1 mentions)
2 protocols using mouse anti p62 antibody
1
Western Blot Analysis of Cellular Stress Markers
2
Western Blot Analysis of Autophagy
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Flag was detected using an anti-Flag M2 HRP antibody (1:2000) (Sigma-Aldrich, St. Louis, MO). The phosphospecific Beclin 1 S90 antibody was produced by PhosphoSolutions (Aurora, CO). Briefly, synthetic peptides corresponding to phosphorylated and dephosphorylated S90 of Beclin 1 were injected to two rabbits and sera were purified using a phosphopeptide affinity column followed by a dephosphopeptide affinity column. The phosphospecific Beclin 1 S90 antibody was used at a concentration of 1:500. Beclin 1, HSP27, p-HSP27, Actin, MK2, p62, LC3, TOM20, PDI, and GAPDH were detected using a rabbit or goat anti-Beclin 1 antibody (Santa Cruz Biotechnology, Dallas, TX, 1:1000 dilution), a rabbit anti-HSP27 antibody (Santa Cruz Biotechnology, 1:200 dilution), a rabbit anti-p-HSP27 antibody (Santa Cruz Biotechnology, 1:200 dilution), an anti-β-Actin HRP antibody (Santa Cruz Biotechnology, 1:2000 dilution), a rabbit anti-MK2 antibody (Cell Signaling Technology, Beverly, MA; 1:1000 dilution), a mouse anti-p62 antibody (Abnova, Walnut, CA; 1:2000 dilution) a rabbit anti-LC3 antibody (Novus Biologicals, Littleton, CO; 1:1000 dilution), a rabbit anti-TOM20 antibody (Santa Cruz Biotechnology; 1:1000 dilution), a rabbit anti-PDI antibody (Cell Signaling Techology; 1:1000 dilution), and a mouse anti-GAPDH antibody (Chemicon International, Temecula, CA; 1:000 dilution).
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