Propargyl-PEG3-bromide

Overview

• Rapid and simple procedure to generate digested peptides
• Simultaneous digestion, purification and concentration at once
• Peptide generation is complete, with no generation of additional artifacts being detected in mass spectrometry
• Peptides are ready for applications such as mass spectrometry and SDS-PAGE
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

This kit is highly efficient in the enzymatic digestion of simple and complex protein samples using trypsin and the subsequent purification of the resulting peptides using a convenient spin column format.  Trypsin is added to the protein sample and bound to the column.  Salts are washed away and the trypsin is then activated to digest proteins.  Peptides are then eluted in a small volume and ready for downstream analysis. The peptides generated are complete, with no additional artifacts being detected in mass spectrometry.  Fifteen micrograms of protein can be processed, digested and purified with each spin column with about 20 minutes of hands-on time (plus trypsin incubation).   The simultaneous protein digestion and volumetric concentration of the purified peptides makes the kit a convenient method for preparing peptides to be analyzed by many downstream applications such as mass spectrometry and more.

Details

Supporting Data

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Kit Specifications
Maximum Protein Input	15 μg
Minimum Protein Input	2 μg
Minimum Elution Volume	30 μL
Time to Process 10 Purifications	20 minutes (Plus a 1-hour incubation)

Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C. This kit is stable for 2 years after the date of shipment.

Component	Cat. 17500 (25 preps)
Wash Solution C	30 mL
Binding Buffer A	4 mL
Column Activation Buffer	3 mL
Micro Spin Columns	25
Collection Tubes	25
Elution tubes (1.7 mL)	25
Product Insert	1

Bis-Sulfone-PEG9-DBCO is a bis-alkylating labeling reagent that is selective for the cysteine sulfur atoms from a native disulfide. These reagents undergo bis-alkylation to conjugate both thiols derived from the two cysteine residues of a reduced native disulfide bond such as the interchain disulfide bonds of an antibody. The reaction results in covalent rebridging of the disulfide bond via a three carbon bridge leaving the protein structurally intact. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Bis-Sulfone-PEG9-DBCO is a bis-alkylating labeling reagent that is selective for the cysteine sulfur atoms from a native disulfide. These reagents undergo bis-alkylation to conjugate both thiols derived from the two cysteine residues of a reduced native disulfide bond such as the interchain disulfide bonds of an antibody. The reaction results in covalent rebridging of the disulfide bond via a three carbon bridge leaving the protein structurally intact. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.

Recently, NGS became a powerful tool to identify the DNA methylation status at the whole genome level with single-base resolution. However, it is well known that bisulfite treatment of the NGS libraries causes tremendous damage to the libraries.

Bisulfite-Seq kit comparison

In the case of the regular bisulfite seq library preparation (library prep before bisulfite conversion), the DNA shearing equipment and the expensive methylated adaptors are required. In addition, the subsequent bisulfite conversion causes tremendous DNA damage to the constructed libraries.

BioDynami has developed a unique library prep technology to solve the problems. The technology uses bisulfite treated DNA as input to avoid the significant library loss caused by bisulfite conversion. Furthermore, DNA shearing step and expensive methylated adaptors are not required with our kit. The DNA polymerase in the kit has high-fidelity amplification ability and uracil tolerance which is ideal for amplification of bisulfite sequencing libraries. The final library is strand specific.

Bisulfite Sequencing Library Prep Kit Workflow

Three index types are available for the kit:

Non-index (Cat.# 30091): Libraries do not have index.

Index (Cat.# 30092): Each primer contains a unique barcode sequence of 6 bases to identify the individual library. Library multiplexing capacity is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30093): The multiplexing of bisulfite sequencing library is up to 96 samples with unique dual indexes. We used a Four-Base Difference Index System to generate indexes that have at least 4 bases different from each other in the 8-base index. The index primers remove NGS errors including index cross-contamination, index hopping, reads mis-assignment etc. Index information can be downloaded here.

Kit advantages

• • • Easy and Quick protocol
      • Easy: Hands-on time only 10 minutes
      • Quick: Total protocol time around 1.5 hours
    • Simple workflow: Less steps
    • Directional library
    • Save magnetic beads more than 50%
    • Guaranteed high Bisulfite sequencing library conversion efficiency
    • Bisulfite treated DNA as input: From 50 ng to 500 ng

The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.