Total protein from hippocampal tissues was extracted using radio- immunoprecipitation assay with phenylmethanesulfonyl fluoride and the protein concentration was determined using the BCA protein assay kit. A total of 40 μg proteins from each sample were loaded on 10% sodium dodecyl sulfate polyacrylamide gels and the target proteins were transferred to polyvinylidene difluoride membranes. In immunoblotting, membranes were blocked with 5% non-fat milk in Tris buffered saline, with Tween-20 (TBST) at room temperature for 1 h and incubated with Atg-5 antibody (1:400, BA3525-2, BOSTER Biological Technology CO. LTD., Huhan, China), light chain 3B (LC3B) antibody (1:500, BM4827, BOSTER), Beclin-1 antibody (1:400, BA3123-2, BOSTER), or β-actin antibody (1: 1000, sc-47778, Santa Cruz Biotechnology, Santa Cruz, CA, United States) at 4°C overnight. Then the membranes were incubated with a horseradish peroxidase-conjugated anti-rabbit or anti-mouse secondary antibody (1:5000, Beyotime) at 37°C for 45 min. Bands were visualized with enhanced chemiluminescence (7 Sea Pharmtech, Shanghai, China) and analyzed on a gel-pro-analyzer (Media Cybernetics, Bethesda, MD, United States).
Ba3123 2
Manufactured by Boster Bio
Sourced in China
The BA3123-2 is a laboratory centrifuge designed for general-purpose use in research and clinical settings. It features a fixed-angle rotor that can accommodate a variety of sample tubes and microplates. The centrifuge operates at a maximum speed of 6,000 rpm and has a maximum relative centrifugal force of 4,000 x g.
Automatically generated - may contain errors
Lab products found in correlation
Bm4827, by Boster Bio (1 mentions)
Sc 47778, by Santa Cruz Biotechnology (1 mentions)
Ba3525 2, by Boster Bio (1 mentions)
Horseradish peroxidase conjugated anti rabbit or anti mouse secondary antibody, by Beyotime (1 mentions)
Gel pro analyzer, by Media Cybernetics (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Chemiluminescence reagent, by Merck Group (1 mentions)
Ba2849, by Boster Bio (1 mentions)
Ab192890, by Abcam (1 mentions)
Ba0325, by Boster Bio (1 mentions)
Ab8245, by Abcam (1 mentions)
3 protocols using ba3123 2
1
Quantifying Autophagy Proteins in Hippocampal Tissue
2
Necroptosis and Autophagy Markers in Colo320 Cells
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Necroptosis and autophagy relative protein markers including RIP-1 (Zinc metalloprotease Rip1), ATG5 (autophagy protein 5) and Beclin-1 were analyzed by western blot in colo320 intestinal tumor cell treated with BF + GCV or BF-rTK + GCV. The antibodies of RIP-1 (BA0346-2) and Beclin-1 (BA3123-2) were purchased from Boster (Wuhan, China) and the antibodies of ATG5 (10181-2-AP) were purchased from Proteintech (Wuhan, China).
3
Protein Expression Analysis of Corneal Tissue
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The ground corneal tissue and processed HCECs were added to NP-40 buffer containing 1 mM PMSF (Genebase, Vancouver, Canada) and incubated on ice for 30 min. After high-speed centrifugation, the supernatants (total proteins) were collected, and the protein concentration in each supernatant was determined using a PierceTM BCA Protein Assay Kit (Cat. No. 23227). Next, a 40 μg sample of total protein from each supernatant was denatured and separated by 10% SDS-PAGE; after which, the protein bands were transferred onto PVDF membranes (Millipore, Burlington, MA, USA), which were subsequently blocked. The membranes were then incubated overnight at 4°C with primary antibodies, followed by a 1 h incubation with a secondary antibody (anti-rabbit IgG, ab6721, 1:15000; Abcam). Finally, the immunostained protein bands were detected with chemiluminescence reagent (Millipore), and photographed using X-ray film. The primary antibodies used were Col1A1 (BA0325, 1:800; Boster), FN (A00564-1, 1:1000; Boster), LC3B (ab192890, 1:2000; Abcam), Beclin1 (BA3123-2, 1:1500; Boster), P62 (BA2849, 1:1000; Boster), and GAPDH (ab8245, 1:5000; Abcam).
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