Pepsin activity is measured using ~2% hemoglobin (pH 2.0), with enzyme diluted to 0.01–0.05 mg/mL in 10 mM HCl and incubated for 10 min at 37°C. Please see the link below to review the entire protocol:
https://www.sigmaaldrich.com/US/en/technical-documents/protocol/protein-biology/enzyme-activity-assays/enzymatic-assay-of-pepsin
P7000
Pepsin from porcine gastric mucosa
powder, ≥250 units/mg solid
Akční nabídka
Synonyma:
Pepsin A, Pepsin from hog stomach
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| Vám/SKU | Dostupnost | Cena |
|---|---|---|
25 g | Obraťte se na zákaznický servis a vyžádejte si informaci o dostupnosti. | 2 800,00 Kč |
100 g | Obraťte se na zákaznický servis a vyžádejte si informaci o dostupnosti. | 8 110,00 Kč |
1 kg | Obraťte se na zákaznický servis a vyžádejte si informaci o dostupnosti. | 45 200,00 Kč |
O této položce
Číslo CAS:
UNSPSC Code:
12352204
eCl@ss:
42010127
EC Number:
232-629-3
NACRES:
NA.54
MDL number:
Specific activity:
≥250 units/mg solid
Biological source:
Porcine gastric mucosa
2 800,00 Kč
Ušetřete až 15 %
Obraťte se na zákaznický servis a vyžádejte si informaci o dostupnosti.
Technický servis
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Porcine gastric mucosa
Quality Segment
form
powder
specific activity
≥250 units/mg solid
mol wt
35 kDa
UniProt accession no.
application(s)
diagnostic assay manufacturing
shipped in
wet ice
storage temp.
2-8°C
Gene Information
pig ... LOC396892(396892)
Application
Pepsin cleavage can be used to produce F(ab′)2 fragments of antibodies. pepsin at www.sigma-aldrich.com/enzymeexplorer.
Pepsin is a peptidase used to digest proteins and is commonly used in the preparation of Fab fragments from antibodies. Pepsin, from porcine gastric mucosa, has been used to hydrolyze dry cervical samples in mice. Product P7000 is provided as a powder and has been used to treat epithelial cells from a single feline mammary carcinoma.
The enzyme from Sigma has been used to simulate in vitro gastric digestion of cooked cod. It has been used to simulate in vitro gastric digestion of cocoa mass and supplemented dietary fiber. It has also been used to increase the fraction of extractable soluble collagen and to lower the immunogenicity of the resulting collagen from bovine dermal tissue.
Biochem/physiol Actions
The enzyme does not cleave at valine, alanine, or glycine linkages. Z-L-tyrosyl-L-phenylalanine, Z-L-glutamyl-L-tyrosine, and Z-L-methionyl-L-tyrosine may be used as substrates for pepsin digestion. Pepsin is inhibited by several phenylalanine-containing peptides.
Preferential cleavage: hydrophobic and aromatic residues in P1 and P1′ postitions. Cleaves Phe-Val, Gln-His, Glu-Ala, Ala-Leu, Leu-Tyr, Tyr-Leu, Gly-Phe, Phe-Phe and Phe-Tyr bonds in the β chain of insulin
Pepsin hydrolyzes peptide bonds, not amide or ester linkages. Pepsin cleaves peptides with an aromatic acid on either side of the peptide bond. Sulfur-containing amino acids increase susceptibility to hydrolysis when they are close to the peptide bond. Pepsin preferentially cleaves at the carboxyl side of phenylalanine and leucine and at the carboxyl side of glutamic acid residues. Cleaves Phe-Val, Gln-His, Glu-Ala, Ala-Leu, Leu-Tyr, Tyr-Leu, Gly-Phe, Phe-Phe and Phe-Tyr bonds in the β chain of insulin
Pepsin is the major proteolytic enzyme produced in the stomach. It digests proteins through the cleavage of interior peptide linkages..
Pepsin is the major proteolytic enzyme produced in the stomach. It digests proteins through the cleavage of interior peptide linkages..
Analysis Note
Optimum pH is 2-4. Active in 4 M urea and 3 M guanidine HCl. Stable at 60 °C. Pepsin is irreversibly inactivated at pH 8.0 - 8.5.
Other Notes
One unit will produce a ΔA280 of 0.001 per min at pH2.0 at 37 °C, measured as TCA-soluble products using hemoglobin as substrate. (Final volume = 16ml. Light path = 1cm.)
View more information on pepsin at www.sigma-aldrich.com/enzymeexplorer.
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Tato položka | |||
|---|---|---|---|
| description powder, ≥250 units/mg solid | description lyophilized powder, ≥3,200 units/mg protein | description powder, slightly beige, 1200-2400 U/mg | description tested according to Ph. Eur. |
| biological source Porcine gastric mucosa | biological source Porcine gastric mucosa | biological source - | biological source - |
| Gene Information pig ... LOC396892(396892) | Gene Information pig ... LOC396892(396892) | Gene Information pig ... LOC396892(396892) | Gene Information pig ... LOC396892(396892) |
| specific activity ≥250 units/mg solid | specific activity ≥3,200 units/mg protein | specific activity 1200-2400 U/mg | specific activity - |
| form powder | form lyophilized powder | form powder | form powder |
| application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing | application(s) - | application(s) - |
| mol wt 35 kDa | mol wt 35 kDa | mol wt 35 kDa | mol wt 35 kDa |
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signalword
Danger
hcodes
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
target_organs
Respiratory system
Skladovací třída
11 - Combustible Solids
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
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9001-75-6
CAS No.
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Could you please provide further details on the method used to determine pepsin activity, including the hemoglobin concentration used as the substrate, the concentration and preparation conditions of the enzyme solution, and the incubation time at 37°C ？
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Is there a favoured solvent to dissolve Pepsin from Porcine Gastric Mucosa (P7000).
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The recommended solubility of this product is 1 mg/mL in 0.01N HCl. Optimum pH is 2-4. Active in 4 M urea and 3 M guanidine HCl. This is stable at 60 °C. Pepsin is irreversibly inactivated at pH 8.0 - 8.5. Enzyme solution (pepsin) – Prepare a 1 mg/mL stock solution in cold (2-8°C) 10 mM HCl. If insoluble material is present, place the stock solution on ice until dissolved. If the pepsin has dissolved, or if insoluble material is still present, after 1 hour, further dilute the 1 mg/mL stock solution to 0.01–0.05 mg/mL with cold 10 mM HCl solution.
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How long can the enzyme be stored and mantain its activity?
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This product does not contain an expiration date. It contains a recommended retest date of roughly two years after the quality release date. A recommended retest date is the period of time during which the product is expected to remain within established stability specifications, provided that it has been stored under defined conditions. After the Retest Date, product samples should be examined to ensure that the product is still in compliance with the established specifications. For more information you may access the "Product Dating Information" document under "ADDITIONAL USEFUL DOCUMENTS ABOUT OUR PRODUCTS" at the bottom of the Quality Services page with this link: https://www.sigmaaldrich.com/US/en/life-science/quality-and-regulatory-management/quality-services.
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Can I freeze stock solutions of Product P7000, Pepsin from porcine gastric mucosa (Pepsin A)?
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Solutions of the enzyme at pH 4.4 should be stable at -20 °C for 2-3 months.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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I've seen the term "1:10000" used to define Product P7000, Pepsin from porcine gastric mucosa (Pepsin A). What does that mean?
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A historical measurement of pepsin activity was the ratio of pepsin to coagulated egg white that the pepsin would digest. Thus 1:10,000 would refer to one part pepsin to 10,000 parts coagulated egg white. More recent analyses use hemoglobin as a substrate, in place of coagulated egg white.
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Product P7000, Pepsin from porcine gastric mucosa (Pepsin A), can be stored at room temperature, but other pepsins are stored in the cooler or freezer. Why is this?
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This relates to autodigestion of pepsin. P7000 is a relatively "crude" pepsin, where other proteins that are present would be more likely digestion targets. With more purified pepsin, the risk of autodigestion increases, and storage at colder temperatures reduces that risk.
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