Proteins were extracted into RIPA buffer (CWBIO, Beijing, China), separated in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and then transferred to 0.22 μm NC membranes (Millipore, MA, USA). The membranes were blocked and probed with antibodies against TOP2α, β-actin, and the anti-rabbit IgG, HRP-linked antibody (Proteintech Group, Inc., Taiwan, China). The bands were detected with BeyoECL Plus (Beyotime Institute of Biotechnology, Shanghai, China) or Super ECL Star (US Everbright Inc., Suzhou, China). Image Lab Software (Bio-Rad, Hercules, CA, USA) was used to analyze the band intensity.
Super ecl star
Manufactured by US Everbright
Sourced in China, United States, Germany
The Super ECL Star is a high-performance chemiluminescent substrate for the detection of horseradish peroxidase (HRP)-labeled proteins in Western blotting applications. It provides a sensitive and stable signal for the visualization of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Image lab software, by Bio-Rad (1 mentions)
0.22 μm nc membranes, by Merck Group (1 mentions)
Beyoecl plus, by Beyotime (1 mentions)
Ripa buffer, by CWBIO (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Capricorn (1 mentions)
Vegfa, by Proteintech (1 mentions)
Hrp conjugated affinipure goat anti rabbit igg h l, by Proteintech (1 mentions)
Hrp conjugated affinipure goat anti mouse igg h l, by Proteintech (1 mentions)
Nuclear protein extraction kit, by Beyotime (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Uvp chemstudio imaging system, by Analytik Jena (1 mentions)
4 protocols using super ecl star
1
Protein Extraction and Western Blot
2
In Vitro Angiogenesis Assay Protocol
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RPMI-1640 Medium 1640 basic (1×) (Gibco by Thermo Fisher Scientific TM, Cat No: C11875500BT), Penicillin-Streptomycin Solution (HyClone, South Logan, UT, USA. Cat No: SV30010), In vitro Angiogenesis Assay Kit (Germany Millipore, Billerica, MA, USA. Cat No: ECM625), Fetal Bovine Serum (FBS, Capricorn scientific, Australia, Cat No: FBS-52A), 0.25% Trypsin-EDTA (Gibco by Life Technologies. Cat No: 25200072), Phosphate Buffered Saline (PBS, HyClone, South Logan, UT, USA. Cat No: SH30256.01), Transwell®Permeable Supports, 6.5 mm Insert, 24 well plate. 8.0 µm polycarbonate Membrance. Tissue Culuture Tramed polystyrene (CORNING. Cat No: 113), VEGFA (GenScript. Cat No: Z02689-10.), β actin Mouse Monoclonal antibody (Proteintech. Cat No: 66009-1-lg), VEGFA Rabbit Polyclonal antibody (Cat No: 66828-1-Ig), VEGFR2 Rabbit Polyclonal antibody (Proteintech. Cat No: 26415-1-AP), MMP2 Rabbit Polyclonal antibody (Proteintech. Cat No: 10373-2-AP), MMP9 Rabbit Polyclonal antibody (Proteintech. Cat No: 10375-2-AP), HRP-conjugated Affinipure Goat Anti-Mouse IgG (H+L) (Proteintech. Cat No: SA00001-1), HRP-conjugated Affinipure Goat Anti-Rabbit IgG (H+L) (Proteintech. Cat No: SA00001-2), Super ECL Star (US EVERBRIGHT. INC, Cat No: S6010-A-100 mL; S6010-B-100 mL), BBD (Xiamen traditional Chinese medicine Co., Ltd. Lot: 150930).
3
Verification of Chicken Serum Antibody Assays
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The GRA7-, OWP8-, and CCp5A-ELISAs results of chicken serum samples were verified by Western blot as previously described [17 (link)]. Briefly, the recombinant proteins were resolved in a 12% SDS-PAGE gel and transferred to PVDF membrane. After an overnight blocking TBST-5% skimmed milk, the membranes were cut into strips and incubated with the serum samples. The membrane strips were further incubated with anti-pig IgG /anti-chicken IgY peroxidase-labeled conjugate antibodies (Thermo Scientific, USA) diluted at 1:5000. After washing, the strips were incubated with Super ECL Star (US Everbright® Inc) and the protein bands were visualized the specific antibodies. A sample was considered positive if the protein bands were observed.
4
Western Blot Analysis of Radiation-Treated Cells
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TSCC cells were seeded in six-well plates at a density of 1 × 105 cells/ml for attaching overnight. Then, cells were treated with IR or PL alone or PL (pretreated for 1 hr) + IR. After 24 hr, cells were washed with PBS and lysed with RIPA buffer. Nuclear proteins were extracted using the nuclear protein extraction kit (Beyotime Co., China). Protein concentrations were measured by using the BCA protein assay kit (Beyotime Co., China). Equal amounts of protein samples were electrophoresed on 8–10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins were transferred onto an immobilon PVDF membrane at 100 V for 2 hr at 4°C. Membranes were probed with primary antibodies overnight at 4°C and then blotted with secondary antibodies for 2 hr. Super ECL Star (US Everbright, Inc.) and the UVP ChemStudio Imaging System (Analytik Jena, Germany) were employed to examine the electrochemiluminescence of indicated proteins.
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