After dewaxing, the sections were subjected to antigen retrieval by high-pressure heating and then exposed to 3% H2O2 for 10 min. After blocking, the sections were incubated with primary antibody for Lin28A (1:100; rabbit, Abcam, ab279647) or Lin28B (1:100; rabbit, Abcam, ab191881) overnight at 4°C, followed by incubation with goat anti-rat IgG (1:200; Abcam) at 37°C for 20 min. After incubation with diaminobenzidine (DAB), the sections were counterstained with hematoxylin. The slides were evaluated by light microscopy (Nikon Eclipse E100).
Ab279647
Manufactured by Abcam
Sourced in United States
Ab279647 is a recombinant monoclonal antibody targeting Cytochrome c oxidase subunit 4 isoform 1. The product is intended for use in Western Blotting applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab191881, by Abcam (2 mentions)
Ab8245, by Abcam (2 mentions)
Eclipse e100, by Nikon (1 mentions)
Goat anti rat igg, by Abcam (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Enhanced chemi luminescence (ecl), by Merck Group (1 mentions)
Ba1051, by Boster Bio (1 mentions)
Ab19857, by Abcam (1 mentions)
Ab8978, by Abcam (1 mentions)
Ab271286, by Abcam (1 mentions)
Ab222782, by Abcam (1 mentions)
Ab133615, by Abcam (1 mentions)
Ab231303, by Abcam (1 mentions)
Ab275377, by Abcam (1 mentions)
Ab189524, by Abcam (1 mentions)
Ab16663, by Abcam (1 mentions)
Ba1054, by Boster Bio (1 mentions)
Ab32072, by Abcam (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
3 protocols using ab279647
1
Immunohistochemical Detection of Lin28A/B
2
Protein Isolation and Western Blotting
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Total protein was isolated using RIPA buffer (P0013E, Beyotime, Shanghai, China), separated by electrophoresis, and transferred onto PVDF membranes (Millipore). The blots were incubated overnight at 4°C with primary antibodies for Lin28A (1: 1000; Abcam, ab279647), Lin28B (1: 2000; Abcam, ab191881), CD63 (1: 1000; Abcam, ab271286), ALIX (1: 1000; Abcam, ab275377), Calhexin-CNX (1: 1000; Abcam, ab133615), CD44 (1: 1000; Abcam, ab189524), CD133 (1: 2000; Abcam, ab222782), OCT4 (1: 1000; Abcam, ab19857), ALCAM (1: 1000; Abcam, ab279580), Vimentin (1: 1000; Abcam, ab8978), E-Cadherin (1: 1000; Abcam, ab231303), CyclinD1 (1: 200; Abcam, ab16663), C-Myc (1: 1000; Abcam, ab32072), and GAPDH (1: 10,000; Abcam, ab8245). After washing, the blots were incubated with secondary antibody HRP Goat anti-Rabbit IgG (1: 1000; BA1054, BOSTER, Wuhan, China) or HRP Goat anti-Mouse IgG (1: 1000; BA1051, BOSTER, Wuhan, China) for 1 h, and the staining was visualized by enhanced chemiluminescence (ECL, Millipore). The blots were cut prior to hybridization with antibodies during blotting and original blots with membrane edges visible were shown in Supplementary files.
3
Protein Extraction and Western Blot Analysis
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The total protein was extracted from brain tissues and HT-22 cells using RIPA lysis buffer (Beyotime Biotechnology, Shanghai, China). Each protein extract (20 μg) was loaded into 12% denatured polyacrylamide gel, followed by transferring into polyvinylidene fluoride membranes (Millipore Co., Ltd., Bedford, MA, USA) for electrophoresis. The membranes were incubated with 5% skim milk at 25°C for 1 h and then with primary antibodies at 4°C overnight. After washing, the membranes were re-incubated with anti-mouse/rabbit secondary antibody of goat IgG H&L (HRP) (ab279647, 1 : 1000, Abcam; ab205718, 1 : 2000, Abcam) at room temperature for 1 h. The immunoreactive bands were detected using the enhanced chemiluminescence system (Amersham Biosciences, Piscataway, NJ, USA) and an electrophoretic image analyser (Bio-Rad, Hemel Hampstead, UK) with GAPDH as the internal reference. The experiment of each group was performed in triplicate. Primary antibodies were used as follows: rabbit monoclonal antibody GPX4 (ab125066, 1 : 1000, Abcam, Cambridge, MA, USA), Lin28A (ab125066, 1 : 1000, Abcam) and mouse monoclonal antibody GAPDH (ab8245, 1 : 500, Abcam).
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