Log In Sign up
The largest database of trusted experimental protocols

Trypsinization

Manufactured by Promega
Visit Supplier

Trypsinization is a laboratory technique used to detach adherent cells from a growth surface. Trypsin, a proteolytic enzyme, is used to break down the cellular adhesions, allowing the cells to be harvested and transferred to a new culture vessel. This process is a essential step in cell culture and maintenance.

Automatically generated - may contain errors

Lab products found in correlation

Endopeptidase lys c, by Fujifilm (2 mentions) Fusion lumos, by Thermo Fisher Scientific (2 mentions) Easylc 1200, by Thermo Fisher Scientific (2 mentions) Ltq orbitrap velos, by Thermo Fisher Scientific (1 mentions) Itraq kit, by AB Sciex (1 mentions)

4 protocols using trypsinization

1

Proteomic Quantification of Protein Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Competitively eluted (3X FLAG peptide) samples, in 1% Triton, were
diluted 2-fold followed by precipitation overnight in 6 volumes ice cold
acetone. Precipitates were dissolved and chemically reduced in 35uL 8M Urea/70mM
ammonium bicarbionate/20mM Dithiothreitol followed by alkylation (50mM
iodoacetamide). Samples were diluted and digested using Endopeptidase LysC (Wako
Chemicals) followed by additional dilution and trypsinization (Promega).
Acidified tryptic peptides were desalted53 (link) and analyzed using nano-LC-MS/MS (EasyLC1200 and Fusion
Lumos operated in High-High mode, ThermoFisher). Data were queried against
UniProt human database (March 2016) concatenated with common contaminants and
quantitated using MaxQuant v. 1.6.0.13 54 (link). False discovery rates of 2% and 1% was applied to
peptide and protein identification. The iBAQ55 (link) values obtained from MaxQuant, were filtered, using
Perseus software56 (link), and the
following filters; 80% of replicates must contain a valid value in either the
‘experiment’ (n=6) and/or ‘control’ (n=2) groups,
protein must be matched to a minimum of 3 razor/unique peptides. Missing values
in the ‘control’ samples were imputed (Perseus) from a normal
distribution. For visualization only, a t-test was performed (Fig. 3g).
Bayraktar E.C., La K., Karpman K., Unlu G., Ozerdem C., Ritter D.J., Alwaseem H., Molina H., Hoffmann H.H., Millner A., Atilla-Gokcumen G.E., Gamazon E.R., Rushing A.R., Knapik E.W., Basu S, & Birsoy K. (2020). Metabolic co-essentiality mapping identifies c12orf49 as a regulator of SREBP processing and cholesterol metabolism. Nature metabolism, 2(6), 487-498.
+ Open protocol
+ Expand
2

Quantitative Proteomic Analysis of Mouse Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols
Samples were reduced using dithiothreitol and alkylated with iodoacetamide followed by overnight trypsinization (Promega, Stockholm, Sweden). Peptides were iTRAQ labelled, pooled and separated by immobilized pH gradient—isoelectric focusing (IPG-IEF) on narrow range pH 3.7–4.9 and 4.00–4.25 strips, as described previously (21 (link)). Extracted fractions from the IPG-IEF were separated using an Agilent 1200 nano-LC system coupled to Thermo Scientific LTQ Orbitrap Velos. Proteome discoverer 1.3 with Sequest-percolator was used to search the UniProt mouse (120 524) database for protein identification, limited to a false discovery rate of <1%.
Roberts T.C., Johansson H.J., McClorey G., Godfrey C., Blomberg K.E., Coursindel T., Gait M.J., Smith C.I., Lehtiö J., EL Andaloussi S, & Wood M.J. (2015). Multi-level omics analysis in a murine model of dystrophin loss and therapeutic restoration. Human Molecular Genetics, 24(23), 6756-6768.
+ Open protocol
+ Expand
3

Proteomic Quantification of Protein Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Competitively eluted (3X FLAG peptide) samples, in 1% Triton, were
diluted 2-fold followed by precipitation overnight in 6 volumes ice cold
acetone. Precipitates were dissolved and chemically reduced in 35uL 8M Urea/70mM
ammonium bicarbionate/20mM Dithiothreitol followed by alkylation (50mM
iodoacetamide). Samples were diluted and digested using Endopeptidase LysC (Wako
Chemicals) followed by additional dilution and trypsinization (Promega).
Acidified tryptic peptides were desalted53 (link) and analyzed using nano-LC-MS/MS (EasyLC1200 and Fusion
Lumos operated in High-High mode, ThermoFisher). Data were queried against
UniProt human database (March 2016) concatenated with common contaminants and
quantitated using MaxQuant v. 1.6.0.13 54 (link). False discovery rates of 2% and 1% was applied to
peptide and protein identification. The iBAQ55 (link) values obtained from MaxQuant, were filtered, using
Perseus software56 (link), and the
following filters; 80% of replicates must contain a valid value in either the
‘experiment’ (n=6) and/or ‘control’ (n=2) groups,
protein must be matched to a minimum of 3 razor/unique peptides. Missing values
in the ‘control’ samples were imputed (Perseus) from a normal
distribution. For visualization only, a t-test was performed (Fig. 3g).
Bayraktar E.C., La K., Karpman K., Unlu G., Ozerdem C., Ritter D.J., Alwaseem H., Molina H., Hoffmann H.H., Millner A., Atilla-Gokcumen G.E., Gamazon E.R., Rushing A.R., Knapik E.W., Basu S, & Birsoy K. (2020). Metabolic co-essentiality mapping identifies c12orf49 as a regulator of SREBP processing and cholesterol metabolism. Nature metabolism, 2(6), 487-498.
+ Open protocol
+ Expand
4

Proteomic Analysis of Oligoasthenoteratozoospermic Sperm

Check if the same lab product or an alternative is used in the 5 most similar protocols
We prepared the samples according to previously reported methods [11 (link)]. We randomly selected two sperm samples from the oligoasthenoteratozoospermic men and two control samples from the healthy subjects. Sperm were dissolved in a 7-M urea, 2-M thiourea, 65-mM DTT, and 1% (v/v) protease inhibitor mixture. Total protein (300 µg) from each sample was processed by reduction, alkylation, and trypsinization (Promega, Madison, WI). The digested samples were reconstituted in 50 µL of 0.5-M triethylammonium bicarbonate. Labeling was performed using an iTRAQ kit per the manufacturer’s protocol (AB Sciex, Foster City, California, USA) [12 (link)]. The sperm samples of NCs were labeled with iTRAQ tags 114 and 115, whereas those of patients with ICSI were labeled with tags 117 and 119. The labeled samples were dried in a rotary vacuum concentrator before further analysis.
Liang J., Zheng Y., Zeng W., Chen L., Yang S., Du P., Wang Y., Yu X, & Zhang X. (2021). Proteomic Profile of Sperm in Infertile Males Reveals Changes in Metabolic Pathways. The Protein Journal, 40(6), 929-939.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!