About 100 mg samples of longissimus thoracis muscle were dissolved in 50% methanol solution at 4°C for 30 min and centrifuged at 10, 000 × g for 10 min, then the supernatant was filtered through glass wool for further analysis (Li et al., 2015 (link)). Serum for free amino acid analysis was pretreatment as previously described (Kong et al., 2009 (link)). Briefly, 1 mL of serum and 2.5 mL of 7.5% trichloracetic acid were mixed thoroughly, then centrifuged at 12, 000 × g at 4°C for 15 min and the supernatant was filtered through a 0.45 μm membrane. Then a volume of 40 μL supernatant for each samples were labeled with iTRAQ reagents (AA 45/32 kit; Applied Biosystems) as recommended by the manufacturer, and analyzed on an Applied Biosystems 3200 Q TRAP LC/MS/MS system equipped with a RP-C18 column.
Itraq reagents aa 45 32 kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The iTRAQ reagents (AA 45/32 kit) are a set of isobaric labeling reagents used in quantitative proteomics analysis. The reagents enable the simultaneous identification and relative quantification of proteins from up to eight different samples. The core function of the iTRAQ reagents is to facilitate the comparative analysis of protein expression levels across multiple samples.
Automatically generated - may contain errors
3 protocols using itraq reagents aa 45 32 kit
1
Muscle and Serum Pretreatment for Amino Acid Analysis
2
Breast Muscle Free Amino Acid Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The contents of free amino acid in breast muscle were analyzed by an amino acid analyzer (L-8900, Hitachi, Japan) as described in the previous study (16 (link)). Approximately 100 mg samples were dissolved using the water–methanol (1: 1, v/v) at 4°C for 30 min and centrifuged at 10 000 × g for 10 min. The supernatant was then filtered through glass wool and stored at −80°C until analysis. After centrifugation, soluble material was separated from insoluble material, and 40 ml of the supernatant was labeled with iTRAQ reagents (AA 45/32 kit; Applied Biosystems, Foster City, CA, USA) as recommended by the manufacturer and analyzed using an Applied Biosystems 3200 Q TRAP LC/MS/MS system equipped with an RP-C18 column (length of 150 mm, diameter of 4.6 mm and particle size of 5 mm). Amino acids in the samples were quantified based on known standards (Shimadzu Oceania, Pickering Laboratories, Mountain View, CA, USA) and their retention times using LC Solution ver. 1.22 SP1 software (Shimadzu, Kyoto, Japan).
3
Quantification of Free Amino Acids in Muscle
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Free amino acid profile was determined in the longissimus dorsi muscle. About 100 mg samples were dissolved in water with methanol (1:1) at 48C for 30 min and centrifuged at 10 000 g for 10 min, then the supernatant was filtered through glass wool and stored at 2 808C until analyses (28) . After centrifugation to separate soluble from insoluble material, 40 ml of the supernatant were labelled with iTRAQ reagents (AA 45/32 kit; Applied Biosystems) as recommended by the manufacturer, and analysed on an Applied Biosystems 3200 Q TRAP LC/MS/MS system equipped with a RP-C18 column (length 150 mm, diameter 4•6 mm and particle size 5 mm) (29) .
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!