To monitor NSC proliferation, we analyzed NSCs by immunostaining of mouse anti–Ki-67 antibody (BD Pharmingen) and Click-IT EdU detection (Invitrogen) after 4 h exposure to EdU, and co-stained with rabbit anti-Sox2 antibody (EMD Millipore) or DAPI (Sigma) to count all cells. Cell images were obtained on AF6000 (Leica) and BZ-X (Keyence). Cell counting was performed using the ImageJ software for images captured on the AF6000, and Hybrid cell count (Keyence), an algorithm for cell counting, for images captured on the BZ-X. Six to eight images were collected under each condition. Inhibitors used for cell culture were obtained from the indicated suppliers: bafilomycin A1 (Sigma), SAR405 (Cayman Chemical), Dynasore (Adipogen), thapsigargin (Adipogen), nocodazole (Adipogen), DAPT (Calbiochem), AG1478 (Cell Signaling Technology), Torin-1 (Cayman Chemical), and rapamycin (Wako). To measure protein stability, cells were incubated with 10 µg/ml cycloheximide (Sigma) to block protein synthesis.
Nocodazole
Manufactured by Adipogen
Nocodazole is a laboratory reagent that acts as a microtubule-depolymerizing agent. It disrupts the formation of the mitotic spindle during cell division, leading to cell cycle arrest at the G2/M phase. Nocodazole is commonly used in cell biology research to synchronize cells and study the dynamics of microtubules and the cell cycle.
Automatically generated - may contain errors
Lab products found in correlation
Af6000, by Leica (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Bafilomycin a1, by Merck Group (1 mentions)
Click it edu detection, by Thermo Fisher Scientific (1 mentions)
Rapamycin, by Fujifilm (1 mentions)
Dynasore, by Adipogen (1 mentions)
Torin 1, by Cayman Chemical (1 mentions)
Ag1478, by Cell Signaling Technology (1 mentions)
Mouse anti ki67 antibody, by BD (1 mentions)
Sar 405, by Cayman Chemical (1 mentions)
Hybrid cell count, by Keyence (1 mentions)
Thapsigargin, by Adipogen (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Prolong diamond antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
Hygromycin b, by Thermo Fisher Scientific (1 mentions)
Polybrene infection transfection reagent, by Merck Group (1 mentions)
Thymidine, by Chem-Impex (1 mentions)
2 protocols using nocodazole
1
Quantifying Neural Stem Cell Proliferation
2
Comprehensive Cell Signaling Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Lipofectamine 2000, Invitrogen (11668019); Lipofectamine RNAiMAX, Invitrogen (13778150); Polybrene Infection/Transfection Reagent, Sigma-Aldrich (TR-1003); GFP-Trap magnetic agarose, ChromoTek (gtma-20); Trypsin Gold Mass Spectrometry Grade, Promega (V5280); Protein G Sepharose, BioVision (6511); Thymidine, Chem-Impex (00306); Propidium iodide, Sigma-Aldrich (P4170); RNase A, Research Products International (R21750); Nocodazole, AdipoGen (AG-CR1-0019); (+)-S-Trityl-L-cysteine, Alfa Aesar (L14384); Puromycin dihydrochloride, Sigma-Aldrich (P8833); Blasticidin S hydrochloride, 10 mg/ml in HEPES buffer, Alfa Aesar (J67216); Hygromycin B, 50 mg/mL in PBS, Invitrogen (10687010); cOmplete Protease Inhibitor Cocktail, Roche (5056489001); Sodium β-glycerophosphate pentahydrate, Alfa Aesar (L03425); Sodium fluoride, Chem-Impex (01523); Sodium pyrophosphate decahydrate, Fisher (S390); Sodium orthovanadate, MP Bio (0215966410); UBE1, R&D Systems (E-304); UBE2C, R&D Systems (E2-654); UBE2S, R&D Systems (E2-690); securin, Abcam (ab87664); D-biotin, Chem-Impex (00033); Streptavidin Alexa Fluor 568, Invitrogen (S11226); Pierce™ High Capacity Streptavidin Agarose, Thermo Fisher (20359); Streptavidin-conjugated HRP, GeneTex (GTX85912); ProLong™ Diamond Antifade Mountant with DAPI, Thermo Fisher (P36971); Clarity™ Western ECL Substrate, Bio-Rad (1705061).
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