Proteins extracted from BEAS-2B cells and HUVECs were measured using a bicinchoninic acid kit (Beyotime Biotechnology, China). Then, the proteins were resolved on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (10%) and transferred to polyvinylidene fluoride (PVDF) membranes (Millipore, MA, USA). The PVDF membranes were incubated using 5% skimmed milk, and then with primary antibodies at 4 °C overnight. Blots were probed using the following antibodies: anti-IL-1β (1: 1, 000, ab234437; Abcam, Cambridge, UK), anti-TNF-α (1: 1, 000, ab183218; Abcam), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; 1: 2, 000, bs0755R; Bioss, China), with GAPDH being the endogenous control. Then, membranes were further incubated for 1 h using a secondary antibody (1:2, 000, b-0311P-HRP; Bioss).
Bs 0755r
Manufactured by Bioss Antibodies
Sourced in China
The Bs-0755R is a laboratory equipment product offered by Bioss Antibodies. It functions as a tool for various research and analysis applications. The core function of this equipment is to provide a reliable and consistent platform for the intended experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh, by Bioss Antibodies (2 mentions)
Bs 0295g, by Bioss Antibodies (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Ab183218, by Abcam (1 mentions)
Ab234437, by Abcam (1 mentions)
Anti gapdh, by Bioss Antibodies (1 mentions)
B 0311p hrp, by Bioss Antibodies (1 mentions)
Bicinchoninic acid kit, by Beyotime (1 mentions)
Ab196495, by Abcam (1 mentions)
Ab192890, by Abcam (1 mentions)
Ab109012, by Abcam (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Antibodies against beclin 1, by Cell Signaling Technology (1 mentions)
Tmz hy 17364, by MedChemExpress (1 mentions)
Proliferating cell nuclear antigen (pcna), by Cell Signaling Technology (1 mentions)
Bcl xl, by Cell Signaling Technology (1 mentions)
Goat anti rabbit secondary antibody, by Biosharp (1 mentions)
Lc3bi 2, by Cell Signaling Technology (1 mentions)
Bs 0295g hrp, by Bioss Antibodies (1 mentions)
D085075, by Bio-Rad (1 mentions)
6 protocols using bs 0755r
1
Western Blot Analysis of Inflammatory Markers
2
Glioblastoma Therapeutic Evaluation Protocol
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2-BP (18,263-25-7) was purchased from Merck (Darmstadt, Germany). TMZ (HY-17364) was purchased from MedChemExpress (New Jersey, USA). Antibodies against P62 (1:1,000; ab109012), LC3B (1:1,000; ab192890), and Bcl2 (1:1,000; ab196495) were purchased from Abcam (Cambridgeshire, England). Antibodies against Beclin-1 (1:1,000; no. 3495), PCNA (1:1,000; no. 13110), Bcl-xl (1:1,000; no. 2764), and caspase-3 (1:1,000; no. 9662) were purchased from Cell Signaling Technology (Massachusetts, USA). Antibodies against GAPDH (1:1,000; bs-0755R) were purchased from Bioss (Beijing, China). A goat anti-rabbit secondary antibody (1:10,000; lot: 70,100,200) was purchased from Biosharp (Wuhan, China).
3
Hippocampal Protein Extraction and Western Blot Analysis
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The damaged hippocampal region tissues were separated with RIPA buffer (Beyotime, China) on ice and homogenized to extract protein. Forty μg proteins were separated with 12% SDS-PAGE (Bio-Rad, China) and transferred to PVDF membranes (EMD Millipore). After blocking with 5% milk for 1.5 h, the membranes were incubated with appropriate primary antibodies diluted with 5% BSA for overnight at 4 °C. The primary antibodies consisted of phospho-PPARγ (ser273) (1:1000, bs-4888R, Bioss, China), LC3BI/II (1:1000, 4108, Cell signaling technology, China), phospho-NF-κB p65 (1:800, bs-230303R, Bioss, China), GAPDH (1:1000, bs-0755R, Bioss, China). After washing with TBS-0.01% Tween 20 for 3 times (10 min/wash), the secondary antibody Goat Anti-rabbit lgG/HRP (1:1000, bs-0295G-HRP, Bioss, China) was cultured with the membranes for 2 h at 25 °C. After washing, the signals were visualized using enhanced chemiluminescence reagent (D085075, Bio-Rad, China).
4
Protein Expression Analysis by Western Blotting
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Tissues or cell lysates were harvested, and the protein concentrations were detected. Then the proteins were separated using 10% SDS-polyacrylamide gel electrophoresis, transferred onto polyvinylidend difluoride membranes, and blocked and incubated with primary antibodies. Finally, the membranes were incubated with the secondary antibody and visualized by enhanced chemiluminescence (SuperSignal Pierce Biotechnology). The antibodies for western blotting were as follows: CD73 (ab133582, Abcam), p-JNK (#4668, Cell Signaling technology), EGFR (ab52894, Abcam) and GAPDH (bs-0755R, Bioss).
5
Cardiac Fibroblast Protein Analysis
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Total protein was extracted from cardiac fibroblasts and myocardial samples. The proteins were separated and transferred to polyvinylidene fluoride (PVDF) membranes. Then, the PVDF membranes were blocked for 2 h at room temperature and incubated overnight at 4°C with one of the following primary antibodies: anti-collagen II (Abcam, ab188570, UK), anti-SMA (Abcam, ab7817, UK), PI3K (Abcam, ab191606, UK), p-PI3K (Abcam, ab182651, UK), AKT (Abcam, ab179463, UK), p-AKT (Abcam, ab192623, UK), mTOR (Abcam, ab134903, UK), p-mTOR (CST, 5536, USA), GAPDH (Bioss, bs-0755 R, China), goat anti-mouse (Bioss, bs-0296 G, China), or goat anti-rabbit (Bioss, bs-0295 G, China). The procedures were performed as previously described, and protein expression was examined by a chemiluminescence analyzer (Azure, A300, USA).
6
Western Blot Analysis of Apoptosis Markers
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Using RIPA lysis buffer (RIPA; Beyotime Institute of Biotechnology) total protein was extracted from the cells. Using bicinchoninic acid (BCA) method (Pierce; Thermo Fisher Scientific, Inc.) the protein was quantified, and each sample (2 µg) was loaded on 10% gels for SDS-PAGE. The protein was transferred on a polyvinylidene difluoride membrane (Millipore Sigma). Then the membranes were blocked with 5% skimmed milk for ~2 h at room temperature and the membrane was soaked in primary antibodies against pro Caspase-3 (1:1,000; cat. no. ab32150; Abcam), cleaved Caspase-3 (1:500; cat. no. ab32042; Abcam) and GAPDH (1:1,000; cat. no. bs-0755R; BIOSS) at 4˚C for 12 h and secondary antibodies (goat anti-rabbit IgG H&L; 1:1,000; cat. no. bs-0295G; BIOSS) for 2 h at room temperature. Bands were visualized by enhanced chemiluminescence (Amersham ECL; cat. no. RPN3243; Cytiva) and analyzed by ImageJ Software v1.53 (National Institutes of Health).
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