To induce ER stress, 100 files with different genotypes were transferred to vials containing 1% agar, 5% sucrose and 12 μM tunicamycin (Sangon Biotech, China) as previously reported. The flies were kept in normal conditions and counted every 3 hours.
Tunicamycin
Manufactured by Sangon
Sourced in China, United States
Tunicamycin is a laboratory reagent commonly used in cell biology research. It is a small molecule that inhibits the first step of protein N-glycosylation, a post-translational modification process essential for the proper folding and function of many proteins. Tunicamycin is often utilized as a tool to study the effects of impaired glycosylation on cellular processes and protein function.
Automatically generated - may contain errors
Lab products found in correlation
Dnase 1, by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by MP Biomedicals (1 mentions)
Phenylmethylsulfonyl fluoride pmsf, by Beyotime (1 mentions)
P mtor ser2448, by Cell Signaling Technology (1 mentions)
P p70s6k, by Cell Signaling Technology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Cytochrome c, by Proteintech (1 mentions)
Caspase 12, by Proteintech (1 mentions)
4e bp1, by Cell Signaling Technology (1 mentions)
P 4ebp1, by Cell Signaling Technology (1 mentions)
P70s6k, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
β actin, by Proteintech (1 mentions)
Caspase 3, by Proteintech (1 mentions)
Bcl2 associated x (bax), by Proteintech (1 mentions)
Bcl 2, by Proteintech (1 mentions)
Thapsigargin, by Thermo Fisher Scientific (1 mentions)
D2650, by Merck Group (1 mentions)
Platinum taq dna polymerase, by Thermo Fisher Scientific (1 mentions)
Goscript reverse transcription system, by Promega (1 mentions)
5 protocols using tunicamycin
1
Inducing ER Stress in Flies
2
PMSF-Based Protein Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phenylmethylsulfonyl fluoride (PMSF) (Cat# ST505) was purchased from Beyotime Biotechnology, Shanghai, China. Lysis buffer containing protease inhibitor (Cat# CW233) was obtained from Cwbio, Beijing, China. The 4-Phenylbutyric acid (Cat# HY-A0281) was purchased from MCE, New Jersey, USA, while tunicamycin (CAS# 11089-65-9) was from Sangon Biotech, Shanghai, China. Dimethyl sulfoxide (CAS#67-68-5) was purchased from MP Biomedicals, Irvine, CA, USA.
3
Dissecting the Unfolded Protein Response Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We bought primary antibodies of GRP78 (Cat#48119), CHOP (SAB 40744) from Signalway Antibody (USA). The primary antibodies of XBP1 (Cat#24868-1-AP), BCL2 (Cat#12789-1-AP), BAX (Cat#50599-2-IG), Caspase3 (Cat#19677-1-AP), Caspase12 (Cat#24868-1-AP), Cytochrome C (Cat# 66264-1-IG), were β-Actin (Cat# 66009-1-IG) were supplied by Proteintech (China). We purchased the primary antibodies from Cell Signaling Technology (USA), and the antibodies include cleaved Caspase3 (Cat#9661s), Caspase3 (Cat#14220S) mTOR (Cat#2983s), p-mTOR ser2448 (Cat#5536s), 4E-BP1 (Cat#9644), p-4E-BP1 (Cat#2855), P70S6K (Cat#9202), and p-P70S6K (Cat#9205). ATF6 (Cat#WL02407) was from Wanleibio (China). We performed the study by using secondary antibodies of HRP-conjugated Goat anti-Rabbit IgG (Cat#HS101-01) and HRP-conjugated Goat anti-mouse IgG (Cat#HS201-01). These antibodies were supplied by TransGEN Biotech (China). Sangon Biotech (China) provided us with Tunicamycin (Cat#A611129-0005).
4
Endoplasmic Reticulum Stress Induction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following reagents were used: Dimethyl sulfoxide (DMSO, D2650, Sigma-Aldrich, Saint Louis, MO, USA), tunicamycin (A611129, Sangon Biotech, Shanghai, China), thapsigargin (T7458, Life Technologies, Carlsbad, CA, USA). Unless otherwise indicated, a final concentration of 2 μg/ml tunicamycin or thapsigargin was used to treat hepatoma cells. DMSO was used as a vehicle control.
5
Quantifying XBP1 Splicing Upon PCV2 Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Activation of IRE1 was determined by measuring splicing of its substrate, the mRNA encoding the XBP1 transcription factor. PK-15 cells were infected with PCV2 for 12 to 36 h. Cells treated with 2 μg/mL tunicamycin (Sangon, Shanghai, China) for 3 h were used as positive control. RNA was harvested using TRIzol reagent (Invitrogen). Total RNA was treated with DNase I (Thermo, Marina, CA, USA) before synthesis of cDNA by reverse transcriptase (The GoScript™ Reverse Transcription System, Promega, Madison, WI, USA). To amplify xbp1 mRNA, PCR was performed for 30 cycles (94 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min (10 min in the final cycle)), using the primers 5'-GGCAGAGACCAAGGGGAATG-3' and 5'-GGGTCGACTTCTGGGAGCTG-3' with Platinum Taq DNA polymerase (Invitrogen). Fragments of 235 bp and 263 bp, representing spliced (sXBP1) and unspliced XBP1 (uXBP1), respectively, were documented after staining the 2% agarose gel with ethidium bromide.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!