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Cat. No.: PPAG-1 (for 1ml)

Cat. No.: PPAG-5 (for 5ml)

Cat. No.: PPAG-20 (for 20ml)

Description

Pepsin Agarose (Pepsin Agarose Gel), also known as Immobilized Pepsin, Pepsin Agarose Resin, or Immobilized Pepsin Agarose Resin, is made by covalently coupling high-quality pepsin to highly cross-linked 6% agarose. This product is highly effective for generating and purifying F(ab')2 fragments from immunoglobulin (IgG) without contaminating F(ab')2 with pepsin. Additionally, it can be used to digest peptide or protein samples containing protease cleavage sites and for producing bioactive peptides through digestion.

Pepsin (EC 3.4.23.1), also known as an acid endopeptidase, is an aspartic protease produced by the interaction of pepsinogen with hydrochloric acid in the gastric mucosa. With a molecular weight of approximately 35kDa, it is commonly isolated and purified from porcine gastric mucosa. Pepsin is a nonspecific acid protease with broad substrate specificity, preferentially cleaving peptide bonds at the P1 and P1' positions of aromatic amino acids such as tyrosine, phenylalanine, and tryptophan. If sulfur-containing amino acids are near these aromatic amino acid peptide bonds, they are also easily hydrolyzed by pepsin. Pepsin can only hydrolyze peptide bonds and cannot hydrolyze non-peptide amide bonds or ester bonds. It exhibits high activity in acidic environments, with optimal activity at a pH of around 1.5-2. In neutral or alkaline environments (pH > 6), pepsin denatures and loses its activity.

This product is widely used in protein structure and function research, proteomics research, and can specifically cleave the heavy chain in the antibody hinge region to generate and purify antibody-active fragments such as divalent antigen molecules (F(ab')2 fragments). It can also be used for protease digestion to produce bioactive peptides.

Features

• High Digestive Capacity: This product boasts a strong digestive capacity compared to similar products, enabling rapid digestion of various IgGs in complex samples. Each milliliter of Pepsin Agarose precipitate contains approximately 2-3mg of pepsin and can digest no less than 40mg of IgG.
• High Stability and Specificity: The immobilized pepsin maintains its activity for a longer duration compared to free pepsin, allowing for overnight incubation. It specifically cleaves peptide bonds of aromatic L-amino acids and heavy chains in the antibody hinge region, resulting in high-purity products that are suitable for subsequent analysis such as Western blotting, ELISA, and mass spectrometry. The agarose used in this product is high-quality 6FF as a matrix, which is highly cross-linked 6% agarose with performance similar to Sepharose 6 Fast Flow. This ensures the stability and purity of the agarose gel, effectively preventing non-specific reactions.
• Convenient Use: This product allows for rapid digestion of IgG, with the enzyme digestion reaction being stopped by simply centrifuging to remove the pepsin agarose gel. This eliminates the need for additional column purification methods, such as ion exchange, to separate fragments from the protease.

• Product Content: 50% settled gel in a specific protective buffer
• Agarose Structure: 6% cross-linked agarose
• Average Particle Size: 45-165μm
• Coupled Protein: Pepsin
• Molecular Weight of Protein: ~35kDa
• Protein Concentration: 2-3mg Pepsin per ml settled gel (>6000 units)
• Digestion Capacity: ≥40mg IgG for 1 ml settled gel
• Specificity: Cleaves somewhat nonspecifically at exposed residues, prefers the carboxyl side of aromatic residues, does not cleave at valine, alanine, or glycine
• Application: Antibody fragmentation, primarily cleaving into F(ab')2 fragments

Storage

• Store at -20ºC for one year.
• Store at 4ºC for at least one month.

Precautions

• Before use, thoroughly resuspend the product by inverting several times to mix the agarose gel evenly. Ensure that the mixing is gentle to avoid denaturation of proteins or breaking of the agarose gel.
• This product contains trace amounts of preservatives. Before use, wash the gel three times with TBS or Digestion Buffer to eliminate any potential interference from the preservatives.
• During purification, it is recommended to set up positive and negative control groups.
• After collecting protein samples, complete the purification work as soon as possible and always keep the samples at 4ºC or on ice to slow down protein degradation or denaturation. To effectively inhibit protein degradation, add an appropriate amount of protease inhibitor mix to the protein sample.
• If centrifugation does not completely remove insoluble substances from the protein sample, filter the sample solution using a 0.45μm filter membrane.
• High concentrations of DTT, beta-mercaptoethanol, and guanidine hydrochloride may affect the binding of this product to the ligand.
• This product is intended for scientific research use by professionals only. It is not for clinical diagnosis or treatment, nor for use in food or medicine. Do not store it in a common household setting.
• For your safety and health, please wear a lab coat and disposable gloves when handling.

Only for research and not intended for treatment of humans or animals

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