All paraffin-embedded LUAD tissues collected from 90 suffers were used for TMA construction and IHC [20 (link)]. The TMA sections which IHC staining was performed on were incubated with anti-BUB1B(#4116, Cell Signaling Technology, Beverly, MA, USA) at 1:100. The final staining score was determined by the intensity (range 0–3+: 0, negative; 1+, weak; 2+, intermediate; and 3+, strong) and the percentage of staining cells(range 1+–4+: 1+, < 25%; 2+, 25%–50%; 3+, 50%–75%; and 4+, > 75%). A final score < 4 was reckoned as low expression and the other scores were taken as high expression.
Anti bub1b
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-BUB1B is a laboratory reagent used to detect the BUB1B protein in cell and tissue samples. BUB1B plays a key role in the spindle assembly checkpoint, which ensures proper chromosome segregation during cell division.
Automatically generated - may contain errors
Lab products found in correlation
Anti hk2, by Abcam (1 mentions)
Anti pkm2, by Abcam (1 mentions)
Ab137852, by Abcam (1 mentions)
Anti glut1, by Abcam (1 mentions)
Anti ldha, by Abcam (1 mentions)
Accutase solution, by Merck Group (1 mentions)
Alamar blue solution, by Thermo Fisher Scientific (1 mentions)
Bradford protein assay, by Bio-Rad (1 mentions)
Albumin from bovine serum, by Merck Group (1 mentions)
Alexa fluor 488 annexin 5 dead cell apoptosis kit, by Thermo Fisher Scientific (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
β actin, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Iscript reverse transcription supermix for qrt pcr, by Bio-Rad (1 mentions)
Pageruler plus prestained protein, by Thermo Fisher Scientific (1 mentions)
Anti foxm1, by Abcam (1 mentions)
3 protocols using anti bub1b
1
Immunohistochemical Profiling of BUB1B in LUAD
2
Protein Quantification and Western Blot Analysis
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Relative protein were extracted from tissue samples or cultured cells, and protein concentration was quantified by Lowry’s protein assay (Bio-Rad, Hercules, CA, USA) [19 (link)]. All protein samples were separated by 10% SDS-PAGE and then transferred onto polyvinylidene fluoride (PVDF) membranes.The primary antibodies used in this study were anti-BUB1B(#4116, Cell Signaling Technology, Beverly, MA, USA), anti-ZNF143(#PA5-72,658, Invitrogen, Waltham, MA, USA), anti-GLUT1(#ab115730, Abcam, Cambridge, UK), anti-LDHA(#ab101562, Abcam, Cambridge, UK), anti-PKM2 (#ab137852, Abcam, Cambridge, UK), anti-HK2 (#ab137852, Abcam, Cambridge, UK) and anti-Actin (#23,600-1-AP, Proteintech, China) antibodies. Western blot analysis was analyzed by Image J software.
3
BUB1B and FOXM1 Modulation in Cell Apoptosis
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The following reagents and primary antibodies were used in the present study: DMEM-F12, fetal bovine serum (FBS), Accutase solution (Sigma-Aldrich, Billerica, MA, USA), alamarBlue solution (all from Thermo Fisher Scientific, Waltham, MA, USA), RIPA buffer, phosphatase inhibitor cocktail, protease inhibitor cocktail (all from Sigma-Aldrich), Bradford protein assay (Bio-Rad Laboratories, Hercules, CA, USA), albumin from bovine serum (BSA; Sigma-Aldrich), PageRuler Plus prestained protein (Thermo Fisher Scientific), iScript Reverse Transcription Supermix for qRT-PCR (Bio-Rad Laboratories), BUB1B overexpression lentivirus (pLenti-GIII-CMV-GFP-2A-Puro; BC018739), shBUB1B lentivirus (piLenti-siRNA-GFP, iV002138), shFOXM1 lentivirus (piLenti-siRNA-GFP, iV008091) [all from Applied Biological Materials (abm) Inc., Richmond, BC, Canada], siomycin A (ALX-380-243-MC05; Enzo Life Sciences, Farmingdale, NY, USA), Alexa Fluor® 488 Annexin V/Dead Cell Apoptosis kit (Thermo Fisher Scientific). Anti-BUB1B (rabbit; #4116; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-FOXM1 (rabbit; ab184637; Abcam, Cambridge, MA, USA), and β-actin (A5316; mouse; Sigma-Aldrich) were used.
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