GST fusion protein isotope ([γ 32 P]ATP) kinase labeling kit product manual (Chinese version)
The main purpose
The YIJI GST fusion protein isotope ([γ 32 P]ATP) kinase labeling reagent is a target fusion protein designed to be purified using the isotope ([γ 32 P]ATP) and cAMP-dependent protein kinase K (protein kinase K). Become the authoritative and classic technical method of sensitive probes. The technology has been carefully developed and successfully tested. Recombinant proteins suitable for a variety of GST fusions. It is applied to subsequent FAR WESTERN BLOT imprinting experiments and BLOT OVERLAY BINDING experiments. The product is ready to use, stable performance, convenient operation, sensitive reaction and good repeatability.
technical background
Glutathione-S-transferase (GST) fusion protein technology is widely used in the study of protein-protein interactions, antibody production and purification, and biochemical analysis. The GST-based fusion protein consists of GSTmoietry, a protease excision site, and a known kinase phosphorylation site, and the purified target fusion protein is labeled using an isotope ([γ 32 P]ATP) and a cAMP-dependent protein kinase K (protein kinase K). As a sensitive probe, the binding protein or polypeptide domain is probed and analyzed in a FAR WESTERN BLOT blotting or BLOT OVERLAY BINDING assay. GST fusion protein isotopes ([γ 32 P]ATP) kinase markers are commonly used tools for proteomics research.
product content
YIJI buffer (Reagent A) ml
YIJI Marking Solution (Reagent B) Microliter (user-supplied)
YIJI Reaction Solution (Reagent C) Microliter
YIJI Enzyme Solution (Reagent D) Microliter
Product manual 1 copy
storage method
Stored in a -20 ° C refrigerator; effective guarantee for June
User-supplied
[γ 32 P]ATP isotope: for isotope labeling
1.5 ml centrifuge tube: container for sample reaction operations and storage
Dry thermostat: for labeling incubation
Liquid scintillation counter: for isotope activity determination
Experimental procedure
- Prepare purified GST fusion protein
- Transfer xx μl of YIJI buffer (Reagent A) to a 1.5 ml centrifuge tube
- Add 2.5 μl of user-prepared GST fusion protein (total 10 μg)
- Add xx microliters of YIJI labeling solution (Reagent B) (Note: total isotope is 10 millicuries per milliliter; 5000 curies per millimolar)
- Add xx microliters of YIJI reaction solution (Reagent C)
- Add xx microliters of YIJI Enzyme Solution (Reagent D)
- Mix the gun head up and down
- Incubate in a 30°C dry thermostat for 30 minutes
- Store at -70 ° C or place in an ice tank
- Or use 5 to 10 microliters (10 7 CPM) of labeled protein to continue subsequent FAR WESTERN BLOT blotting and BLOT OVERLAY BINDING experiments.
Precautions
- This product is 20 operations
- The marking efficiency of this product can reach more than 60%
- Wear gloves when handling
- Strict sterility during operation
- Isotope operations must strictly abide by the rules and regulations
- It is recommended to operate with a gun head
- Isotope [γ 32 P]ATP (1 Curie = 37 GBq), ICN or AMERSHAM is recommended.
- Due to the short half-life of the [γ 32 P] isotope, it is recommended to use fresh isotope materials;
- It can be directly labeled on GST fusion protein resin, then purified by conventional elution or thrombin (THROMBIN); isotope activity determination after purification
- The company provides a series of isotope processing reagent products
Quality Standard
- This product has been certified to be stable.
- This product has been identified as free of contaminating proteases and ribozymes.
Organic Alfalfa Grass Powder,Alfalfa Leaf Powder,Organic Alfalfa Leaf Powder,Young Leaf Powder Alfalfa
Hengshui Shanzhi Health Drink Co., Ltd , https://www.grasspowder-sz.com