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A. What is the peptide sequence? How long is the peptide?
Peptides can be from a few amino acids to over 100 amino acids. Peptides over 25 amino acids are more difficult and may require consultation.
B. What is the required purity?
- Polyclonal antibody production requires about 80-85% purity- Post-translational modification specific antibody production (phospho, methyl, acetyl, etc) requires about 85-90% purity - Purities of 90-95%: This is the sweet spot where most people will be happy. Almost all high level work can utilize this purity range. - For non-critical experiments, epitope mapping, and assays that do not require high purity, peptides a purity of about 70% can be used.
C. Are there special modifications, non-natural amino acids, or other requirements?
The most common modifications, referred to as PTMs (post-translational modifications), include phosphorylated serine, threonine and tyrosine, epigenetic modifications such as methylated lysine (mono-, di- or tri-methylated), arginine (mono-, asymmetric- or symmetric dimethylated), and acetylated lysine.
D. How much peptide is required?
For most applications 10 mg is sufficient if the peptide is used for immunizations and 20 mg if the peptide will be used for generation of affinity column matrix for antibody purification. Generation of post-translational modification antibodies requires multiple purification steps and will require more than a single peptide.
Rockland’s TrueBlot® product line enables researchers to generate publication-quality IP western blot data. Simply substitute your conventional HRP blotting reagent with Goat TrueBlot®, Mouse TrueBlot®, Rabbit TrueBlot® or Sheep TrueBlot®.
TrueBlot® is a horseradish peroxidase conjugated immunoblotting detection reagent, which enables unhindered detection of blotted target bands. This results in unparalleled clarity in results and high quality imaging suitable for reproduction.
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