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Merck

T3924

Trypsin-EDTA solution

1 ×, sterile; sterile-filtered, BioReagent, suitable for cell culture, 0.5 g porcine trypsin and 0.2 g EDTA, 4Na per liter of Hanks′ Balanced Salt Solution with phenol red

동의어(들):

Cell dissociation enzyme

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제품정보 (DICE 배송 시 비용 별도)

EC 번호:
UNSPSC Code:
12352204
NACRES:
NA.78
MDL number:
Biological source:
Porcine
Concentration:
1 ×
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도움 문의

biological source

Porcine

sterility

sterile; sterile-filtered

product line

BioReagent

form

solution

mol wt

23.4 kDa

concentration

1 ×

technique(s)

cell culture | mammalian: suitable, single cell analysis: suitable

impurities

Porcine parvovirus, none detected (9 CFR)

pH

7.0-9.0

shipped in

dry ice

storage temp.

−20°C

Quality Level

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Application

The typical use for this product is in removing adherent cells from a culture surface. The concentration of trypsin necessary to dislodge cells from their substrate is dependent primarily on the cell type and the age of the culture.

Suitable for use in preparation of single cell suspension for sequencing.

Biochem/physiol Actions

Trypsin cleaves peptides on the C-terminal side of lysine and arginine residues. The rate of hydrolysis of this reaction is slowed if an acidic residue is on either side of the cleavage site and hydrolysis is stopped if a proline residue is on the carboxyl side of the cleavage site. The optimal pH for trypsin activity is 7-9. Trypsin can also act to cleave ester and amide linkages of synthetic derivatives of amino acids. EDTA is added to trypsin solutions as a chelating agent that neutralizes calcium and magnesium ions that obscure the peptide bonds on which trypsin acts. Removing these ions increases the enzymatic activity.

Serine protease inhibitors, including DFP, TLCK, APMSF, AEBSEF, and aprotinin, amongst others, will inhibit Trypsin.

Preparation Note

This product does contain phenol red. Due to shipment on dry ice, there could be significant carbon dioxide buildup in the package. This CO2 may enter the solution and lower the pH slightly, giving an orange rather than pinkish color. The orange solution will still be suitable for use, or the pH can be adjusted with sodium hydroxide. Incubating cells with too high a trypsin concentration for a long period can damage cell membranes and kill the cells. Solubilizing trypsin or diluting it from a concentrated solution should be done with a buffered salt solution containing no Ca2+ or Mg2+.

Other Notes

Trypsin consists of a single chain polypeptide of 223 amino acid residues, produced by the removal of the N-terminal hexapeptide from trypsinogen which is cleaved at the Lys - lle peptide bond. The sequence of amino acids is cross-linked by 6 disulfide bridges. This is the native form of trypsin, beta-trypsin. BETA-trypsin can be autolyzed, cleaving at the Lys - Ser residue, to produce alpha-trypsin. Trypsin is a member of the serine protease family.

Disclaimer

This product is stored frozen between -10 and -40°C. Repeated cycles of freezing and thawing should be avoided.

저장 등급

12 - Non Combustible Liquids

wgk

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


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문서

Use of MULTI-seq lipid-modified oligos, protocol, and troubleshooting guide for PCR Assays and Sequencing applications.

Technical information for working with human coronary artery smooth muscle cells including thawing, subculturing and cryopreservation

The field of proteomics is continually looking for new ways to investigate protein dynamics within complex biological samples. Recently, many researchers have begun to use RNA interference (RNAi) as a method of manipulating protein levels within their samples, but the ability to accurately determine these protein amounts remains a challenge. Fortunately, over the past decade, the field of proteomics has witnessed significant advances in the area of mass spectrometry. These advances, both in instrumentation and methodology, are providing researchers with sensitive assays for both identification and quantification of proteins within complex samples. This discussion will highlight some of these methodologies, namely the use of Multiple Reaction Monitoring (MRM) and Protein-AQUA.

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프로토콜

Technical information for working with human aortic smooth muscle cells including thawing, subculturing and cryopreservation.

Trypsin is commonly used for dissociating adherent cells from surfaces. A wide variety of trypsin solutions are available to meet your specific cell line requirements.

Cell culture protocol for high cell viability freezing using cryopreservation reagents like DMSO. Free handbook download.

Store the cryovials in a liquid nitrogen storage tank immediately upon arrival.

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