Medaka Sea is one of the most popular models of fish species for Ecotoxicology and environmental research and study of proteomics is a useful tool for understanding the medaka molecular response after exposure to different environmental stresses. High-quality protein sample preparation is the key to producing high-quality two-dimensional gel electrophoresis (2-DE) results for proteomics analysis.
In recent years, the extraction of protein-based Trizol has gained popularity because of its performance promise in producing high-quality 2-DE and comfort method.Three Trizol-based approach (Trizol method, the method and the method of Trizol Trizol aliquots with commercial cleaning kit) is used to extract protein from marine samples medaka and 2-DE profiles produced. 2-DE profile quality and effectiveness of extraction methods were evaluated.
For comparison, two common protein extraction method (method of lysis buffer and trichloroacetic acid (TCA) / acetone precipitation extraction) are also applied in parallel to Trizol based approaches.Any three Trizol-based approach produces high-quality 2-DE profiles of marine medaka compared with both methods of lysis buffer and TCA / acetone precipitation extraction.
In addition, the Trizol method with clean-up kit produced the best commercial 2-DE profile in terms of the clarity of the background, the number of spots and resolution proteins.Trizol based approach offered a better choice than traditional protein extraction method for 2-DE analysis of medaka sea. A modified version of the method Trizol with commercial clean-up kit was shown to produce the best 2-DE profile.
Production of high-quality two-dimensional gel electrophoresis profile for marine medaka samples by using Trizol-based protein extraction approaches.
A method of combining blue native polyacrylamide gel electrophoresis liquid chromatography-tandem mass spectrometry for the detection of circulating immune complexes between the monoclonal antibody therapy and anti-drug antibodies in animals.
Monoclonal antibody therapy can potentially induce unwanted immune response, so that the production of anti-drug antibodies (ADAS). Binding of fennel drug and subsequent formation of immune complexes (ICs) can trigger a variety of responses, depending on the size, concentration, and a subclass of fennel. To better understand the impact of fennel in pharmacokinetics, pharmacodynamics and toxicology profile, bioanalytical methods developed to detect IC between a human monoclonal immunoglobulin G (IgG) and fennel in a biological sample.
Regarding the experimental procedure, in brief, human-specific antibodies and human antibodies bound IC in biological samples separated by blue native polyacrylamide gel electrophoresis (BN-PAGE). The target fraction is then cut from the gel, followed by in-gel trypsin digestion and subsequent liquid chromatography-tandem mass spectrometry (LC-MS / MS) to monitor the human peptide-specific IgG. This method is capable of detecting various types of human antibodies with a lower detection limit of 10 ug / mL in monkey serum. Performance IC assay to detect the expressed using spiked samples, and pre-incubated in monkey serum IC clearly detectable.
Description: A polyclonal antibody for detection of GELS from Human, Mouse, Rat. This GELS antibody is for WB, ELISA. It is affinity-purified from rabbit serum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from part region of human GELS protein
Description: A polyclonal antibody for detection of GELS from Human, Mouse, Rat. This GELS antibody is for WB, ELISA. It is affinity-purified from rabbit serum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from part region of human GELS protein
Description: A polyclonal antibody for detection of GELS from Human, Mouse, Rat. This GELS antibody is for WB, ELISA. It is affinity-purified from rabbit serum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from part region of human GELS protein
AXYGEN® GEL TRAY 10 X 10 CM FOR USE WITH 10 CM GEL BOX, UV TRANSPARENT
Description: Km: 0.89 mM for neuraminidase X-Neu5Ac is a new substrate for chromogenic assay of neuraminidase activity in bacterial expression systems.
10 Well Comb ,1mm Thick For Enduro 15cm Gel System
Description: 10-DEBC hydrochloride is a selective inhibitor of Akt (or termed PKB) [1], with an IC50 value of approximate 48 ?M [2].Akt is a type of serine/threonine kinase. It phosphorylates and inactivates components in the apoptotic machinery, including Caspase 9 and BAD.
Description: AI-10-49 is a selective inhibitor of CBF? -SMMHC and RUNX1 interaction with a FRET IC50 value of 260nM. AI-10-49 restores RUNX1 transcriptional activity, displays favorable pharmacokinetics, and delays leukemia progression in mice.
Description: HG-10-102-01 is a potent and selective inhibitor of leucine-rich repeat kinase 2 (LRRK2) with the IC50 values of 20.3nM and 3.2nM for wild type LRRK2 and LRRK2 [G1019S], respectively [1].
Taken together, these findings indicate that our method allows semi-quantitative analysis to estimate the ratio of human antibodies compared with the IC included in the total antibody. This method was successfully applied to the in vivo studies using rats, and the data helps explain the unexpectedly rapid clearance of human antibodies for the formation of large IC. The combination of IC separation by BN-PAGE and detection of peptide-specific human IgG with LC-MS / MS bioanalytical approach is common, useful for detecting IC in animals.
