Chensinin-1b was synthesized as per the standard Fmoc solid-phase peptide synthetic approach by GL Biochem Co., Ltd. (Shanghai, China). Reverse-phase HPLC was used to purify the synthesized peptide to 95% homogeneity by using a 2.2 × 25 cm Vydac 218TP1022 C-18 column (Separations Group, Hesperis, CA, USA) equilibrated with acetonitrile/water/tri-fluoroacetic acid. After purification, a MALDI-TOF mass spectrometer (Shimadzu, Kyoto, Japan) was utilized for the detection of the relative mass of chensinin-1b.
Maldi tof mass spectrometer
Manufactured by Shimadzu
Sourced in Japan
The MALDI-TOF mass spectrometer is an analytical instrument used for the identification and characterization of molecules. It employs the matrix-assisted laser desorption/ionization (MALDI) technique to generate ions from samples, which are then detected and analyzed by a time-of-flight (TOF) mass analyzer. This core function allows for the accurate determination of the molecular masses of various compounds, including proteins, peptides, lipids, and other biomolecules.
Automatically generated - may contain errors
Lab products found in correlation
Anhydrous organic solvents, by Merck Group (2 mentions)
Silica gel 60, by Merck Group (2 mentions)
Drx 400, by Bruker (2 mentions)
Silica gel 60 f254, by Merck Group (1 mentions)
Drx 500, by Bruker (1 mentions)
Scanning electron microscopy, by Zeiss (1 mentions)
Transmission electron microscopy, by JEOL (1 mentions)
Zetasizer 3000 ha, by Malvern Panalytical (1 mentions)
Mascot, by Matrix Science (1 mentions)
Cc 105, by TOMY (1 mentions)
Drx 500 spectrometer, by Bruker (1 mentions)
Yl9100, by Younglin (1 mentions)
Axima performance, by Shimadzu (1 mentions)
8 protocols using maldi tof mass spectrometer
1
Chensinin-1b Peptide Synthesis and Purification
2
Synthesis and Characterization of DSPE-PEG-Based Conjugates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DSPE-PEG2000-COOH (25 mg, 0.0088 mmol) or DSPE-PEG2000-NH2 (25 mg, 0.0088 mmol) was allowed to react for 3 h with EDC-HCl (3.4 mg, 0.018 mmol) and NHS (1.01 mg, 0.0087 mmol) in 2 mL of chloroform in the presence of three drops of triethylamine at room temperature in the dark. The progress of the reaction was checked by thin-layer chromatography (TLC) on a silica gel plate. Next, dequalinium (DQA; 4.6 mg, 0.0087 mmol) or TPP (3.9 mg, 0.0087 mmol) dissolved in DMSO (2 mL) was added to DSPE-PEG2000-COO-NHS and stirred for 30 min. Chloroform was removed by an evaporator, and 5 mL of distilled water was added. The final mixture was dialyzed using a cellulose dialysis tubing (MWCO 3000) against deionized water for 48 h, and then freeze-dried using a lyophilizer. After freeze-drying, the residue was redissolved in distilled water and filtered through a 0.45-µm filter syringe. The liquid was then freeze-dried again and the formed product was characterized by proton NMR spectroscopy (Brucker, 600 MHz, Billerica, MA, USA) and MALDI-TOF mass spectrometer (AXIMA-Assurance, Shimadzu, Kyoto, Japan).
3
Purification and Characterization of Organic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All reagents and starting materials were purchased from commercial chemical suppliers (Sigma-Aldrich, TCI and Across Organics) and used as received. All the anhydrous organic solvents of purity greater than 99.9% were purchased from Aldrich and used directly. Thin layer chromatography (TLC) was performed on Merck aluminum sheets with silica gel 60 F254 and were visualized by ultraviolet light, KMnO4 staining and ninhydrin. For the purification of compounds, column chromatography was performed on Merck silica gel 60 (70–230 mesh or 230–400 mesh). 1H and 13C NMR spectra were recorded on a Bruker DRX-400 and DRX-500 spectrometer. Chemical shifts (δ) are reported in parts per million (ppm) measured relative to an internal standard and coupling constants (J) are expressed in hertz (Hz). Mass spectra were recorded using Shimadzu (MALDI-TOF) mass spectrometer.
4
Nanoparticle Characterization and Stability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Characterization of the nanoparticles was carried out with scanning electron microscopy (Zeiss, Germany) and transmission electron microscopy (JEOL, Japan). The three-dimensional structure was measured by atomic force microscopy (ATM, CU, China). The size, polydispersity index (PDI) and zeta potential were determined by a Nano ZS (Zetasizer 3000 HAS, Malvern Instrument, England). A MALDI-TOF mass spectrometer (Shimadzu, Japan) was used to evaluate the stability of the proteins in the nanoparticles. The entrapment efficiency was calculated according to the following formulas: Entrapment efficiency (%) = (Amounttotal Vo − Amountloading Vo)/Amounttotal Vo × 100.
5
Characterization of PEGylated VEGF
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot assay, Size-exclusion HPLC (SE-HPLC) and MALDI-TOF mass spectroscopy were used for characterization of PEGylated VEGF. PEGylation of VEGF was examined by using Western blot assay. Briefly anti-VEGF rabbit polyclonal antibody (1 : 500 dilution), and goat anti-rabbit antibody conjugated to horseradish peroxidase (HRP) (1 : 5000 dilution) were used as primary and secondary antibody respectively. Also, west pico chemiluminescent substrate was applied for signal generation.
SE-HPLC was performed to identify the different fraction of PEGylated VEGF using BIOshell A400 Protein C4, 2.1 mm × 15 cm column. The diluted sample in PBS buffer (1X, pH 7.4) was injected to the column with a flow rate of 0.5 mL min−1; 280 nm UV detector was used to monitor the elution peaks.
MALDI-TOF mass spectroscopy was performed in linear acquisition operation mode using MALDI TOF Mass Spectrometer (Shimadzu-7090). Briefly, MALDI-target was coated with a saturated solution of sinapinic acid in ethanol. Subsequently, the purified sample (10 µL) was mixed with 10 µL of sinapinic acid (1% in 40% acetonitrile), and 1 µL of the resulting mixture was loaded onto the target.
SE-HPLC was performed to identify the different fraction of PEGylated VEGF using BIOshell A400 Protein C4, 2.1 mm × 15 cm column. The diluted sample in PBS buffer (1X, pH 7.4) was injected to the column with a flow rate of 0.5 mL min−1; 280 nm UV detector was used to monitor the elution peaks.
MALDI-TOF mass spectroscopy was performed in linear acquisition operation mode using MALDI TOF Mass Spectrometer (Shimadzu-7090). Briefly, MALDI-target was coated with a saturated solution of sinapinic acid in ethanol. Subsequently, the purified sample (10 µL) was mixed with 10 µL of sinapinic acid (1% in 40% acetonitrile), and 1 µL of the resulting mixture was loaded onto the target.
6
Proteomic Analysis of LNCaP and AILNCaP Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The spots that exhibited differences between LNCaP and AILNCaP14 or AILNCaP15 cells in image analysis were excised, destained with acetonitrile, reduced with DTT, carbamidemethylated with iodoacetamide, and digested with trypsin (APRO SCIENCE, Tokushima, Japan). The collected solutions were dried using a centrifugal concentrator CC‐105 (TOMY SEIKO Co., LTD., Tokyo, Japan), mixed with 2, 5‐dihydroxybenzoic acid as a matrix, and analyzed using a MALDI‐TOF mass spectrometer (AXIMA Resonance and/or Performance, Shimadzu Corporation, Kyoto, Japan). The data were analyzed using Mascot (Matrix Science Ltd., London, UK). Details of experimental procedures were presented in supporting material and methods (Doc S1 ).
7
Cyanine Dye Synthesis and Purification
8
MALDI-ToF Mass Spectrometry of Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mass spectra were collected using the linear mode of AXIMA Performance (Shimadzu) MALDI-ToF mass spectrometer equipped with an N2 laser (337 nm) at a laser power of 130 W. Protein samples were mixed with 3,5-dimethoxy-4-hydroxycinnamic acid (Sinapinic acid) matrix at volumetric ratios of 1:1 on a stainless-steel target plate and dried thoroughly at room temperature before analysis. Representative spectra were obtained by averaging 200 spectra within the mass range of 3,500-16,000 Da, and presented in smooth gauss mode of 200.
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!