enzymatic  (Millipore)


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    Name:
    Trypsin EDTA solution
    Description:

    Catalog Number:
    t4174
    Price:
    None
    Applications:
    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. Trypsin-EDTA solution is used for the following applications:. Used as a supplement in cell culture for their maintenance. In harvesting cells grown to confluence. to detach lentivirus-transduced macrophages
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    Structured Review

    Millipore enzymatic

    https://www.bioz.com/result/enzymatic/product/Millipore
    Average 99 stars, based on 41 article reviews
    Price from $9.99 to $1999.99
    enzymatic - by Bioz Stars, 2020-12
    99/100 stars

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    Related Articles

    RNA Extraction:

    Article Title: Effects of Therapeutic Doses of Celecoxib on Several Physiological Parameters of Cultured Human Osteoblasts
    Article Snippet: .. RNA extraction and cDNA synthesis (reverse transcription) After 24 h of culture with Celecoxib treatment (untreated cells served as controls), cells were detached from the culture flask using 0.05% trypsin-EDTA solution (Sigma) and individually harvested. mRNA was extracted using a silicate gel technique in the QiagenRNeasy extraction kit (Qiagen Inc., Hilden, Germany), which includes a DNAse digestion step. .. The amount of extracted mRNA was measured by UV spectrophotometry at 260 nm (Eppendorf AG, Hamburg, Germany), and contamination with proteins was determined according to the 260/280 ratio.

    Fluorescence:

    Article Title: The Spalt transcription factors regulate cell proliferation, survival and epithelial integrity downstream of the Decapentaplegic signalling pathway
    Article Snippet: .. Fluorescence Activated Cell Sorting (FACS) We incubated wing imaginal discs of UAS-dicer2/+ ; salEPv -Gal4 UAS-GFP/UAS-salm-i; UAS-salr-i/+ and UASdicer2/+ ; salEPv -Gal4 UAS-GFP/+ (control discs) in 300 µl of trypsin solution (Trypsin–EDTA Sigma T4299) and 0.5 µl Hoescht (Hoescht33342, Trihydrochloride Trihydrate H3570, Molecular ProbesTM ) at 28°C during 40 minutes. ..

    Transfection:

    Article Title: Functional genomics platform for pooled screening and mammalian genetic interaction maps
    Article Snippet: .. Target cell line in which the screen is conducted 293T cells (ATCC cat # CRL-3216) Heat-inactivated FBS (Hyclone Characterized, Thermo Scientific, Cat# SH30071.03) DMEM-High glucose (Sigma, cat# D5796) RPMI w/HEPES modification (Sigma, cat# R5886) 100x Penicillin/Streptomycin (Sigma, cat# P4333) 100x L-glutamine (Sigma, cat# G7513) PBS (Sigma, cat# D8537) Trypsin (Sigma, cat#T3924) Puromycin (Sigma, cat#P8833-10MG) Mirus LT1 Transfection agent (Mirus, cat # MIR 2300) QIAamp DNA Blood Maxi Kit (Qiagen, cat # 51192) Qiagen miniprep kit (Qiagen, cat # 27106) Qiagen Buffer PB (Qiagen, cat # 19066) Qiagen Gel extraction kit (Cat No. 28704 or 28706) Qiagen Minelute kit (Cat # 28204) Qiagen QIAquick PCR purification kit (Cat No. 28104 or 28106) Ethidium bromide (Biorad, cat # 161-0433) BstXI (NEB, cat # R0113S) PvuII (NEB, cat # R0151S) High-sensitivity DNA kit (Agilent Technologies, cat. no. 5067-4626) TruSeq SBS v3 kit, 50 cycles (Illumina, cat. no. FC-401-3002) TruSeq SR Cluster Kit v3, cBot, HS (Illumina, cat. no. GD-401-3001) Phusion® High-Fidelity PCR Kit with 5X HF buffer (NEB Cat No. E0553L) dNTPs (e.g. NEB N0447L) Polybrene (Sigma cat # H9268-5G) DMSO (Sigma Hybrimax, cat# D2650-5X5ML) Mr. Frosty Freezing container (Fisher, cat# 15-350-50) Tris base (Fisher, cat # BP152-5) Boric acid (Sigma, cat # B6768) Acetic acid (Fisher, cat # A38-212) EDTA (Sigma, cat # 03660) Bromophenol blue (Bio-rad, cat # 161-0611) Xylene cyanol (Sigma, cat # X4126) Sucrose (Sigma, cat # 84097) HEPES (Sigma, cat # H3375) Magnesium acetate (Sigma, cat # M0631) Potassium acetate (Sigma, cat # P5708) Agarose (Research Products International, Cat # A20090-500) 20% Polyacrylamide/TBE gels, 1.0mm, 12-well (Invitrogen, cat # EC63152BOX) 1kb+ DNA ladder (Invitrogen, cat # 10787-018) D-tube midi electroelution columns (EMD cat # 71507-3) Milli-Q purified H2 O Oligonucleotides from IDT Technologies (see sequences in ) MegaX DH10 T1R Electrocompetent Cells (Invitrogen, cat # C6400-03) Lentiviral packaging plasmids pRSV, pMDL, pVSVG (obtained from the Jacks lab, described in refs , ) Plasmid pMK1200 (ref. ), which we will provide on request. .. T4 DNA Ligase + buffer (NEB, cat # M0202T) DH5a cells, subcloning efficiency (Life Technologies, cat#18265-017) Carbenicillin (Research Products International, cat # ) agar (Fisher cat # DF0145070) bacto tryptone (Fisher cat # DF0123-07-5) sodium chloride (Fisher cat # S271-10) yeast extract (Fisher cat # DF0127071) SOC (Life Technologies, cat#15544-034) Pasteur pipets (glass) (VWR cat # 14672-380) Qiagen Gigaprep Kit (Qiagen, cat#12191) Qtray plates (Molecular devices, cat # X6023) 1-mm gap cuvette for electroporation (USA Scientific, cat # 9104-1050)

    Cell Culture:

    Article Title: Chitosan/polyethylene glycol fumarate blend films for wound dressing application: in vitro biocompatibility and biodegradability assays
    Article Snippet: .. Cultured mouse fibroblasts (L929, the cell bank of National Institute of Genetic Engineering and Biotechnology, Iran) were harvested with a 0.25% Trypsin–EDTA solution (Sigma) in a phosphate-buffered saline (PBS; pH 7.4) and re-suspended in the culture medium composed of RPMI-1640 with l -glutamine (Sigma, USA), supplemented with 10% fetal bovine serum (FBS, Gibco, Germany), 1% Pen-Strep (100 IU/mL penicillin and 10 mg/mL streptomycin, Sigma-Aldrich, USA). .. A cell suspension was prepared with a concentration of approximately 1 × 106 cell/mL and seeded onto 12-well plates in contact with the sterilized films.

    Article Title: Decellularized human ovarian scaffold based on a sodium lauryl ester sulfate (SLES)-treated protocol, as a natural three-dimensional scaffold for construction of bioengineered ovaries
    Article Snippet: .. The ovarian cells were then harvested by 0.025% Trypsin/EDTA solution (T3924, Sigma-Aldrich), seeded onto the scaffolds by simple pipetting at a density of 2.0 × 106 cells/scaffold and cultured with 400 μL of plating medium on 24-well plates at 37 °C and 5% CO2 for 1 day. .. Viability of ovarian cells was assessed by MTT test on some randomly selected seeded scaffolds before grafting.

    Purification:

    Article Title: Functional genomics platform for pooled screening and mammalian genetic interaction maps
    Article Snippet: .. Target cell line in which the screen is conducted 293T cells (ATCC cat # CRL-3216) Heat-inactivated FBS (Hyclone Characterized, Thermo Scientific, Cat# SH30071.03) DMEM-High glucose (Sigma, cat# D5796) RPMI w/HEPES modification (Sigma, cat# R5886) 100x Penicillin/Streptomycin (Sigma, cat# P4333) 100x L-glutamine (Sigma, cat# G7513) PBS (Sigma, cat# D8537) Trypsin (Sigma, cat#T3924) Puromycin (Sigma, cat#P8833-10MG) Mirus LT1 Transfection agent (Mirus, cat # MIR 2300) QIAamp DNA Blood Maxi Kit (Qiagen, cat # 51192) Qiagen miniprep kit (Qiagen, cat # 27106) Qiagen Buffer PB (Qiagen, cat # 19066) Qiagen Gel extraction kit (Cat No. 28704 or 28706) Qiagen Minelute kit (Cat # 28204) Qiagen QIAquick PCR purification kit (Cat No. 28104 or 28106) Ethidium bromide (Biorad, cat # 161-0433) BstXI (NEB, cat # R0113S) PvuII (NEB, cat # R0151S) High-sensitivity DNA kit (Agilent Technologies, cat. no. 5067-4626) TruSeq SBS v3 kit, 50 cycles (Illumina, cat. no. FC-401-3002) TruSeq SR Cluster Kit v3, cBot, HS (Illumina, cat. no. GD-401-3001) Phusion® High-Fidelity PCR Kit with 5X HF buffer (NEB Cat No. E0553L) dNTPs (e.g. NEB N0447L) Polybrene (Sigma cat # H9268-5G) DMSO (Sigma Hybrimax, cat# D2650-5X5ML) Mr. Frosty Freezing container (Fisher, cat# 15-350-50) Tris base (Fisher, cat # BP152-5) Boric acid (Sigma, cat # B6768) Acetic acid (Fisher, cat # A38-212) EDTA (Sigma, cat # 03660) Bromophenol blue (Bio-rad, cat # 161-0611) Xylene cyanol (Sigma, cat # X4126) Sucrose (Sigma, cat # 84097) HEPES (Sigma, cat # H3375) Magnesium acetate (Sigma, cat # M0631) Potassium acetate (Sigma, cat # P5708) Agarose (Research Products International, Cat # A20090-500) 20% Polyacrylamide/TBE gels, 1.0mm, 12-well (Invitrogen, cat # EC63152BOX) 1kb+ DNA ladder (Invitrogen, cat # 10787-018) D-tube midi electroelution columns (EMD cat # 71507-3) Milli-Q purified H2 O Oligonucleotides from IDT Technologies (see sequences in ) MegaX DH10 T1R Electrocompetent Cells (Invitrogen, cat # C6400-03) Lentiviral packaging plasmids pRSV, pMDL, pVSVG (obtained from the Jacks lab, described in refs , ) Plasmid pMK1200 (ref. ), which we will provide on request. .. T4 DNA Ligase + buffer (NEB, cat # M0202T) DH5a cells, subcloning efficiency (Life Technologies, cat#18265-017) Carbenicillin (Research Products International, cat # ) agar (Fisher cat # DF0145070) bacto tryptone (Fisher cat # DF0123-07-5) sodium chloride (Fisher cat # S271-10) yeast extract (Fisher cat # DF0127071) SOC (Life Technologies, cat#15544-034) Pasteur pipets (glass) (VWR cat # 14672-380) Qiagen Gigaprep Kit (Qiagen, cat#12191) Qtray plates (Molecular devices, cat # X6023) 1-mm gap cuvette for electroporation (USA Scientific, cat # 9104-1050)

    Modification:

    Article Title: Functional genomics platform for pooled screening and mammalian genetic interaction maps
    Article Snippet: .. Target cell line in which the screen is conducted 293T cells (ATCC cat # CRL-3216) Heat-inactivated FBS (Hyclone Characterized, Thermo Scientific, Cat# SH30071.03) DMEM-High glucose (Sigma, cat# D5796) RPMI w/HEPES modification (Sigma, cat# R5886) 100x Penicillin/Streptomycin (Sigma, cat# P4333) 100x L-glutamine (Sigma, cat# G7513) PBS (Sigma, cat# D8537) Trypsin (Sigma, cat#T3924) Puromycin (Sigma, cat#P8833-10MG) Mirus LT1 Transfection agent (Mirus, cat # MIR 2300) QIAamp DNA Blood Maxi Kit (Qiagen, cat # 51192) Qiagen miniprep kit (Qiagen, cat # 27106) Qiagen Buffer PB (Qiagen, cat # 19066) Qiagen Gel extraction kit (Cat No. 28704 or 28706) Qiagen Minelute kit (Cat # 28204) Qiagen QIAquick PCR purification kit (Cat No. 28104 or 28106) Ethidium bromide (Biorad, cat # 161-0433) BstXI (NEB, cat # R0113S) PvuII (NEB, cat # R0151S) High-sensitivity DNA kit (Agilent Technologies, cat. no. 5067-4626) TruSeq SBS v3 kit, 50 cycles (Illumina, cat. no. FC-401-3002) TruSeq SR Cluster Kit v3, cBot, HS (Illumina, cat. no. GD-401-3001) Phusion® High-Fidelity PCR Kit with 5X HF buffer (NEB Cat No. E0553L) dNTPs (e.g. NEB N0447L) Polybrene (Sigma cat # H9268-5G) DMSO (Sigma Hybrimax, cat# D2650-5X5ML) Mr. Frosty Freezing container (Fisher, cat# 15-350-50) Tris base (Fisher, cat # BP152-5) Boric acid (Sigma, cat # B6768) Acetic acid (Fisher, cat # A38-212) EDTA (Sigma, cat # 03660) Bromophenol blue (Bio-rad, cat # 161-0611) Xylene cyanol (Sigma, cat # X4126) Sucrose (Sigma, cat # 84097) HEPES (Sigma, cat # H3375) Magnesium acetate (Sigma, cat # M0631) Potassium acetate (Sigma, cat # P5708) Agarose (Research Products International, Cat # A20090-500) 20% Polyacrylamide/TBE gels, 1.0mm, 12-well (Invitrogen, cat # EC63152BOX) 1kb+ DNA ladder (Invitrogen, cat # 10787-018) D-tube midi electroelution columns (EMD cat # 71507-3) Milli-Q purified H2 O Oligonucleotides from IDT Technologies (see sequences in ) MegaX DH10 T1R Electrocompetent Cells (Invitrogen, cat # C6400-03) Lentiviral packaging plasmids pRSV, pMDL, pVSVG (obtained from the Jacks lab, described in refs , ) Plasmid pMK1200 (ref. ), which we will provide on request. .. T4 DNA Ligase + buffer (NEB, cat # M0202T) DH5a cells, subcloning efficiency (Life Technologies, cat#18265-017) Carbenicillin (Research Products International, cat # ) agar (Fisher cat # DF0145070) bacto tryptone (Fisher cat # DF0123-07-5) sodium chloride (Fisher cat # S271-10) yeast extract (Fisher cat # DF0127071) SOC (Life Technologies, cat#15544-034) Pasteur pipets (glass) (VWR cat # 14672-380) Qiagen Gigaprep Kit (Qiagen, cat#12191) Qtray plates (Molecular devices, cat # X6023) 1-mm gap cuvette for electroporation (USA Scientific, cat # 9104-1050)

    Incubation:

    Article Title: The Spalt transcription factors regulate cell proliferation, survival and epithelial integrity downstream of the Decapentaplegic signalling pathway
    Article Snippet: .. Fluorescence Activated Cell Sorting (FACS) We incubated wing imaginal discs of UAS-dicer2/+ ; salEPv -Gal4 UAS-GFP/UAS-salm-i; UAS-salr-i/+ and UASdicer2/+ ; salEPv -Gal4 UAS-GFP/+ (control discs) in 300 µl of trypsin solution (Trypsin–EDTA Sigma T4299) and 0.5 µl Hoescht (Hoescht33342, Trihydrochloride Trihydrate H3570, Molecular ProbesTM ) at 28°C during 40 minutes. ..

    Article Title: Increased PD-L1 expression in radioresistant HNSCC cell lines after irradiation affects cell proliferation due to inactivation of GSK-3beta
    Article Snippet: .. Cells were detached by incubation with 0,05% trypsin-EDTA solution (Sigma-Aldrich, Munich, Germany) for 5 to 10 minutes (min) at 37°C. ..

    Gel Extraction:

    Article Title: Functional genomics platform for pooled screening and mammalian genetic interaction maps
    Article Snippet: .. Target cell line in which the screen is conducted 293T cells (ATCC cat # CRL-3216) Heat-inactivated FBS (Hyclone Characterized, Thermo Scientific, Cat# SH30071.03) DMEM-High glucose (Sigma, cat# D5796) RPMI w/HEPES modification (Sigma, cat# R5886) 100x Penicillin/Streptomycin (Sigma, cat# P4333) 100x L-glutamine (Sigma, cat# G7513) PBS (Sigma, cat# D8537) Trypsin (Sigma, cat#T3924) Puromycin (Sigma, cat#P8833-10MG) Mirus LT1 Transfection agent (Mirus, cat # MIR 2300) QIAamp DNA Blood Maxi Kit (Qiagen, cat # 51192) Qiagen miniprep kit (Qiagen, cat # 27106) Qiagen Buffer PB (Qiagen, cat # 19066) Qiagen Gel extraction kit (Cat No. 28704 or 28706) Qiagen Minelute kit (Cat # 28204) Qiagen QIAquick PCR purification kit (Cat No. 28104 or 28106) Ethidium bromide (Biorad, cat # 161-0433) BstXI (NEB, cat # R0113S) PvuII (NEB, cat # R0151S) High-sensitivity DNA kit (Agilent Technologies, cat. no. 5067-4626) TruSeq SBS v3 kit, 50 cycles (Illumina, cat. no. FC-401-3002) TruSeq SR Cluster Kit v3, cBot, HS (Illumina, cat. no. GD-401-3001) Phusion® High-Fidelity PCR Kit with 5X HF buffer (NEB Cat No. E0553L) dNTPs (e.g. NEB N0447L) Polybrene (Sigma cat # H9268-5G) DMSO (Sigma Hybrimax, cat# D2650-5X5ML) Mr. Frosty Freezing container (Fisher, cat# 15-350-50) Tris base (Fisher, cat # BP152-5) Boric acid (Sigma, cat # B6768) Acetic acid (Fisher, cat # A38-212) EDTA (Sigma, cat # 03660) Bromophenol blue (Bio-rad, cat # 161-0611) Xylene cyanol (Sigma, cat # X4126) Sucrose (Sigma, cat # 84097) HEPES (Sigma, cat # H3375) Magnesium acetate (Sigma, cat # M0631) Potassium acetate (Sigma, cat # P5708) Agarose (Research Products International, Cat # A20090-500) 20% Polyacrylamide/TBE gels, 1.0mm, 12-well (Invitrogen, cat # EC63152BOX) 1kb+ DNA ladder (Invitrogen, cat # 10787-018) D-tube midi electroelution columns (EMD cat # 71507-3) Milli-Q purified H2 O Oligonucleotides from IDT Technologies (see sequences in ) MegaX DH10 T1R Electrocompetent Cells (Invitrogen, cat # C6400-03) Lentiviral packaging plasmids pRSV, pMDL, pVSVG (obtained from the Jacks lab, described in refs , ) Plasmid pMK1200 (ref. ), which we will provide on request. .. T4 DNA Ligase + buffer (NEB, cat # M0202T) DH5a cells, subcloning efficiency (Life Technologies, cat#18265-017) Carbenicillin (Research Products International, cat # ) agar (Fisher cat # DF0145070) bacto tryptone (Fisher cat # DF0123-07-5) sodium chloride (Fisher cat # S271-10) yeast extract (Fisher cat # DF0127071) SOC (Life Technologies, cat#15544-034) Pasteur pipets (glass) (VWR cat # 14672-380) Qiagen Gigaprep Kit (Qiagen, cat#12191) Qtray plates (Molecular devices, cat # X6023) 1-mm gap cuvette for electroporation (USA Scientific, cat # 9104-1050)

    Polymerase Chain Reaction:

    Article Title: Functional genomics platform for pooled screening and mammalian genetic interaction maps
    Article Snippet: .. Target cell line in which the screen is conducted 293T cells (ATCC cat # CRL-3216) Heat-inactivated FBS (Hyclone Characterized, Thermo Scientific, Cat# SH30071.03) DMEM-High glucose (Sigma, cat# D5796) RPMI w/HEPES modification (Sigma, cat# R5886) 100x Penicillin/Streptomycin (Sigma, cat# P4333) 100x L-glutamine (Sigma, cat# G7513) PBS (Sigma, cat# D8537) Trypsin (Sigma, cat#T3924) Puromycin (Sigma, cat#P8833-10MG) Mirus LT1 Transfection agent (Mirus, cat # MIR 2300) QIAamp DNA Blood Maxi Kit (Qiagen, cat # 51192) Qiagen miniprep kit (Qiagen, cat # 27106) Qiagen Buffer PB (Qiagen, cat # 19066) Qiagen Gel extraction kit (Cat No. 28704 or 28706) Qiagen Minelute kit (Cat # 28204) Qiagen QIAquick PCR purification kit (Cat No. 28104 or 28106) Ethidium bromide (Biorad, cat # 161-0433) BstXI (NEB, cat # R0113S) PvuII (NEB, cat # R0151S) High-sensitivity DNA kit (Agilent Technologies, cat. no. 5067-4626) TruSeq SBS v3 kit, 50 cycles (Illumina, cat. no. FC-401-3002) TruSeq SR Cluster Kit v3, cBot, HS (Illumina, cat. no. GD-401-3001) Phusion® High-Fidelity PCR Kit with 5X HF buffer (NEB Cat No. E0553L) dNTPs (e.g. NEB N0447L) Polybrene (Sigma cat # H9268-5G) DMSO (Sigma Hybrimax, cat# D2650-5X5ML) Mr. Frosty Freezing container (Fisher, cat# 15-350-50) Tris base (Fisher, cat # BP152-5) Boric acid (Sigma, cat # B6768) Acetic acid (Fisher, cat # A38-212) EDTA (Sigma, cat # 03660) Bromophenol blue (Bio-rad, cat # 161-0611) Xylene cyanol (Sigma, cat # X4126) Sucrose (Sigma, cat # 84097) HEPES (Sigma, cat # H3375) Magnesium acetate (Sigma, cat # M0631) Potassium acetate (Sigma, cat # P5708) Agarose (Research Products International, Cat # A20090-500) 20% Polyacrylamide/TBE gels, 1.0mm, 12-well (Invitrogen, cat # EC63152BOX) 1kb+ DNA ladder (Invitrogen, cat # 10787-018) D-tube midi electroelution columns (EMD cat # 71507-3) Milli-Q purified H2 O Oligonucleotides from IDT Technologies (see sequences in ) MegaX DH10 T1R Electrocompetent Cells (Invitrogen, cat # C6400-03) Lentiviral packaging plasmids pRSV, pMDL, pVSVG (obtained from the Jacks lab, described in refs , ) Plasmid pMK1200 (ref. ), which we will provide on request. .. T4 DNA Ligase + buffer (NEB, cat # M0202T) DH5a cells, subcloning efficiency (Life Technologies, cat#18265-017) Carbenicillin (Research Products International, cat # ) agar (Fisher cat # DF0145070) bacto tryptone (Fisher cat # DF0123-07-5) sodium chloride (Fisher cat # S271-10) yeast extract (Fisher cat # DF0127071) SOC (Life Technologies, cat#15544-034) Pasteur pipets (glass) (VWR cat # 14672-380) Qiagen Gigaprep Kit (Qiagen, cat#12191) Qtray plates (Molecular devices, cat # X6023) 1-mm gap cuvette for electroporation (USA Scientific, cat # 9104-1050)

    FACS:

    Article Title: The Spalt transcription factors regulate cell proliferation, survival and epithelial integrity downstream of the Decapentaplegic signalling pathway
    Article Snippet: .. Fluorescence Activated Cell Sorting (FACS) We incubated wing imaginal discs of UAS-dicer2/+ ; salEPv -Gal4 UAS-GFP/UAS-salm-i; UAS-salr-i/+ and UASdicer2/+ ; salEPv -Gal4 UAS-GFP/+ (control discs) in 300 µl of trypsin solution (Trypsin–EDTA Sigma T4299) and 0.5 µl Hoescht (Hoescht33342, Trihydrochloride Trihydrate H3570, Molecular ProbesTM ) at 28°C during 40 minutes. ..

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Functional genomics platform for pooled screening and mammalian genetic interaction maps
    Article Snippet: .. Target cell line in which the screen is conducted 293T cells (ATCC cat # CRL-3216) Heat-inactivated FBS (Hyclone Characterized, Thermo Scientific, Cat# SH30071.03) DMEM-High glucose (Sigma, cat# D5796) RPMI w/HEPES modification (Sigma, cat# R5886) 100x Penicillin/Streptomycin (Sigma, cat# P4333) 100x L-glutamine (Sigma, cat# G7513) PBS (Sigma, cat# D8537) Trypsin (Sigma, catT3924) Puromycin (Sigma, cat#P8833-10MG) Mirus LT1 Transfection agent (Mirus, cat # MIR 2300) QIAamp DNA Blood Maxi Kit (Qiagen, cat # 51192) Qiagen miniprep kit (Qiagen, cat # 27106) Qiagen Buffer PB (Qiagen, cat # 19066) Qiagen Gel extraction kit (Cat No. 28704 or 28706) Qiagen Minelute kit (Cat # 28204) Qiagen QIAquick PCR purification kit (Cat No. 28104 or 28106) Ethidium bromide (Biorad, cat # 161-0433) BstXI (NEB, cat # R0113S) PvuII (NEB, cat # R0151S) High-sensitivity DNA kit (Agilent Technologies, cat. no. 5067-4626) TruSeq SBS v3 kit, 50 cycles (Illumina, cat. no. FC-401-3002) TruSeq SR Cluster Kit v3, cBot, HS (Illumina, cat. no. GD-401-3001) Phusion® High-Fidelity PCR Kit with 5X HF buffer (NEB Cat No. E0553L) dNTPs (e.g. NEB N0447L) Polybrene (Sigma cat # H9268-5G) DMSO (Sigma Hybrimax, cat# D2650-5X5ML) Mr. Frosty Freezing container (Fisher, cat# 15-350-50) Tris base (Fisher, cat # BP152-5) Boric acid (Sigma, cat # B6768) Acetic acid (Fisher, cat # A38-212) EDTA (Sigma, cat # 03660) Bromophenol blue (Bio-rad, cat # 161-0611) Xylene cyanol (Sigma, cat # X4126) Sucrose (Sigma, cat # 84097) HEPES (Sigma, cat # H3375) Magnesium acetate (Sigma, cat # M0631) Potassium acetate (Sigma, cat # P5708) Agarose (Research Products International, Cat # A20090-500) 20% Polyacrylamide/TBE gels, 1.0mm, 12-well (Invitrogen, cat # EC63152BOX) 1kb+ DNA ladder (Invitrogen, cat # 10787-018) D-tube midi electroelution columns (EMD cat # 71507-3) Milli-Q purified H2 O Oligonucleotides from IDT Technologies (see sequences in ) MegaX DH10 T1R Electrocompetent Cells (Invitrogen, cat # C6400-03) Lentiviral packaging plasmids pRSV, pMDL, pVSVG (obtained from the Jacks lab, described in refs , ) Plasmid pMK1200 (ref. ), which we will provide on request. .. T4 DNA Ligase + buffer (NEB, cat # M0202T) DH5a cells, subcloning efficiency (Life Technologies, cat#18265-017) Carbenicillin (Research Products International, cat # ) agar (Fisher cat # DF0145070) bacto tryptone (Fisher cat # DF0123-07-5) sodium chloride (Fisher cat # S271-10) yeast extract (Fisher cat # DF0127071) SOC (Life Technologies, cat#15544-034) Pasteur pipets (glass) (VWR cat # 14672-380) Qiagen Gigaprep Kit (Qiagen, cat#12191) Qtray plates (Molecular devices, cat # X6023) 1-mm gap cuvette for electroporation (USA Scientific, cat # 9104-1050)

    Plasmid Preparation:

    Article Title: Functional genomics platform for pooled screening and mammalian genetic interaction maps
    Article Snippet: .. Target cell line in which the screen is conducted 293T cells (ATCC cat # CRL-3216) Heat-inactivated FBS (Hyclone Characterized, Thermo Scientific, Cat# SH30071.03) DMEM-High glucose (Sigma, cat# D5796) RPMI w/HEPES modification (Sigma, cat# R5886) 100x Penicillin/Streptomycin (Sigma, cat# P4333) 100x L-glutamine (Sigma, cat# G7513) PBS (Sigma, cat# D8537) Trypsin (Sigma, cat#T3924) Puromycin (Sigma, cat#P8833-10MG) Mirus LT1 Transfection agent (Mirus, cat # MIR 2300) QIAamp DNA Blood Maxi Kit (Qiagen, cat # 51192) Qiagen miniprep kit (Qiagen, cat # 27106) Qiagen Buffer PB (Qiagen, cat # 19066) Qiagen Gel extraction kit (Cat No. 28704 or 28706) Qiagen Minelute kit (Cat # 28204) Qiagen QIAquick PCR purification kit (Cat No. 28104 or 28106) Ethidium bromide (Biorad, cat # 161-0433) BstXI (NEB, cat # R0113S) PvuII (NEB, cat # R0151S) High-sensitivity DNA kit (Agilent Technologies, cat. no. 5067-4626) TruSeq SBS v3 kit, 50 cycles (Illumina, cat. no. FC-401-3002) TruSeq SR Cluster Kit v3, cBot, HS (Illumina, cat. no. GD-401-3001) Phusion® High-Fidelity PCR Kit with 5X HF buffer (NEB Cat No. E0553L) dNTPs (e.g. NEB N0447L) Polybrene (Sigma cat # H9268-5G) DMSO (Sigma Hybrimax, cat# D2650-5X5ML) Mr. Frosty Freezing container (Fisher, cat# 15-350-50) Tris base (Fisher, cat # BP152-5) Boric acid (Sigma, cat # B6768) Acetic acid (Fisher, cat # A38-212) EDTA (Sigma, cat # 03660) Bromophenol blue (Bio-rad, cat # 161-0611) Xylene cyanol (Sigma, cat # X4126) Sucrose (Sigma, cat # 84097) HEPES (Sigma, cat # H3375) Magnesium acetate (Sigma, cat # M0631) Potassium acetate (Sigma, cat # P5708) Agarose (Research Products International, Cat # A20090-500) 20% Polyacrylamide/TBE gels, 1.0mm, 12-well (Invitrogen, cat # EC63152BOX) 1kb+ DNA ladder (Invitrogen, cat # 10787-018) D-tube midi electroelution columns (EMD cat # 71507-3) Milli-Q purified H2 O Oligonucleotides from IDT Technologies (see sequences in ) MegaX DH10 T1R Electrocompetent Cells (Invitrogen, cat # C6400-03) Lentiviral packaging plasmids pRSV, pMDL, pVSVG (obtained from the Jacks lab, described in refs , ) Plasmid pMK1200 (ref. ), which we will provide on request. .. T4 DNA Ligase + buffer (NEB, cat # M0202T) DH5a cells, subcloning efficiency (Life Technologies, cat#18265-017) Carbenicillin (Research Products International, cat # ) agar (Fisher cat # DF0145070) bacto tryptone (Fisher cat # DF0123-07-5) sodium chloride (Fisher cat # S271-10) yeast extract (Fisher cat # DF0127071) SOC (Life Technologies, cat#15544-034) Pasteur pipets (glass) (VWR cat # 14672-380) Qiagen Gigaprep Kit (Qiagen, cat#12191) Qtray plates (Molecular devices, cat # X6023) 1-mm gap cuvette for electroporation (USA Scientific, cat # 9104-1050)

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  • 99
    Millipore enzymatic protein deglycosylation kit
    Effect of V-ATPase inhibition on mTORC1/ATF4 pathway in βTC3 cells and processing of Ac45 and PRR in βTC3 (WT and furKO) cells. (A) Western blot for p-S6K, total S6K, p-4E-BP1, total 4E-BP1, ATF4 in βTC3 cells incubated with the V-ATPase inhibitor bafilomycin A1 (50 nM final concentration) for 24h. GAPDH was used as a loading control. (B) Western blot quantification from n=3 independent experiments. (C) Schematic illustration of Ac45 and PRR protein structures. Furin and SKI-1 cleavage sites are indicated by arrows. SP, signal peptide; TM, transmembrane domain; AA, amino acids, SKI-1 (Subtilisin Kexin Isozyme-1). (D) HEK293T and βTC3 (WT and furKO) cells were transfected with a Flag-tagged Ac45 construct. Western blot of HEK293T cell lysate before and after N-glycosidase F (N-glyc F) treatment, with the processed form (24 kDa) appearing only after <t>deglycosylation</t> (left HEK293T panel). Western blot of βTC3 cell lysate, wild type (WT) and furin knockout (furKO), both treated with N-glyc F (right panel). The indicated positions of proAc45 and Ac45 correspond to the predicted MW of the deglycosylated peptide backbone (46 kDa and 24 kDa respectively, black arrows). (E) Full length (fl) and soluble (s)PRR levels in the lysate (upper panel) and conditioned medium (lower panel) of furin knockout (furKO) and WT βTC3 cells. Three independent furKO clones were assessed. Quantifications on the right-hand side show n=3 independent experiments. *P
    Enzymatic Protein Deglycosylation Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/enzymatic protein deglycosylation kit/product/Millipore
    Average 99 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
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    91
    Millipore cathepsin l enzymatic activity
    Comparison of changes in muscle protein synthesis and protein degradation between hindlimb muscles in response to unloading. Adult (9 mo) male rats underwent hindlimb unloading (HU) for 3, 7, or 14 days. Protein synthesis ( A ), 20S and 26S β 5 proteasome subunit activity ( B ), and <t>cathepsin</t> L activity ( C ) were measured in the soleus (open squares), medial gastrocnemius (MG; solid circles), and tibialis anterior (TA; shaded triangles) at each unloading time point. Total protein determined by stain-free imaging of the PVDF membrane was used to normalize puromycin protein expression. All values are expressed as a percentage relative to the control group for each muscle ( n = 5–6/muscle). Values are means ± SE. * P
    Cathepsin L Enzymatic Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cathepsin l enzymatic activity/product/Millipore
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cathepsin l enzymatic activity - by Bioz Stars, 2020-12
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    91
    Millipore caspase 3 enzymatic assay caspase 3 activity
    Cell cycle, apoptotic and proteolytic profiles in MDC1A and Surf1-LS muscle cells. ( A ) Cell distribution in the different phases of the cell cycle. ( B ) Differences in gene expression of CASP3, PSMA2, MURF1, MAFBX and USP19 in MDC1A and Surf1-LS myoblasts and myotubes compared to control. ( C ) <t>Caspase-3</t> enzymatic activity was increased in myotubes from MDC1A and Surf1-LS patients compared to control cells. ( D ) 20S proteasome enzymatic activity was increased in both myoblasts and myotubes from MDC1A patients but was unchanged in Surf1-LS muscle cells compared to control. Results are expressed as a fold change of controls. Columns represent mean values and bars SE. Statistical significance was assessed by one-way ANOVA followed by Duncan’s post hoc test. p
    Caspase 3 Enzymatic Assay Caspase 3 Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/caspase 3 enzymatic assay caspase 3 activity/product/Millipore
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    caspase 3 enzymatic assay caspase 3 activity - by Bioz Stars, 2020-12
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    Image Search Results


    Effect of V-ATPase inhibition on mTORC1/ATF4 pathway in βTC3 cells and processing of Ac45 and PRR in βTC3 (WT and furKO) cells. (A) Western blot for p-S6K, total S6K, p-4E-BP1, total 4E-BP1, ATF4 in βTC3 cells incubated with the V-ATPase inhibitor bafilomycin A1 (50 nM final concentration) for 24h. GAPDH was used as a loading control. (B) Western blot quantification from n=3 independent experiments. (C) Schematic illustration of Ac45 and PRR protein structures. Furin and SKI-1 cleavage sites are indicated by arrows. SP, signal peptide; TM, transmembrane domain; AA, amino acids, SKI-1 (Subtilisin Kexin Isozyme-1). (D) HEK293T and βTC3 (WT and furKO) cells were transfected with a Flag-tagged Ac45 construct. Western blot of HEK293T cell lysate before and after N-glycosidase F (N-glyc F) treatment, with the processed form (24 kDa) appearing only after deglycosylation (left HEK293T panel). Western blot of βTC3 cell lysate, wild type (WT) and furin knockout (furKO), both treated with N-glyc F (right panel). The indicated positions of proAc45 and Ac45 correspond to the predicted MW of the deglycosylated peptide backbone (46 kDa and 24 kDa respectively, black arrows). (E) Full length (fl) and soluble (s)PRR levels in the lysate (upper panel) and conditioned medium (lower panel) of furin knockout (furKO) and WT βTC3 cells. Three independent furKO clones were assessed. Quantifications on the right-hand side show n=3 independent experiments. *P

    Journal: bioRxiv

    Article Title: Furin controls β cell function via mTORC1 signaling

    doi: 10.1101/2020.04.09.027839

    Figure Lengend Snippet: Effect of V-ATPase inhibition on mTORC1/ATF4 pathway in βTC3 cells and processing of Ac45 and PRR in βTC3 (WT and furKO) cells. (A) Western blot for p-S6K, total S6K, p-4E-BP1, total 4E-BP1, ATF4 in βTC3 cells incubated with the V-ATPase inhibitor bafilomycin A1 (50 nM final concentration) for 24h. GAPDH was used as a loading control. (B) Western blot quantification from n=3 independent experiments. (C) Schematic illustration of Ac45 and PRR protein structures. Furin and SKI-1 cleavage sites are indicated by arrows. SP, signal peptide; TM, transmembrane domain; AA, amino acids, SKI-1 (Subtilisin Kexin Isozyme-1). (D) HEK293T and βTC3 (WT and furKO) cells were transfected with a Flag-tagged Ac45 construct. Western blot of HEK293T cell lysate before and after N-glycosidase F (N-glyc F) treatment, with the processed form (24 kDa) appearing only after deglycosylation (left HEK293T panel). Western blot of βTC3 cell lysate, wild type (WT) and furin knockout (furKO), both treated with N-glyc F (right panel). The indicated positions of proAc45 and Ac45 correspond to the predicted MW of the deglycosylated peptide backbone (46 kDa and 24 kDa respectively, black arrows). (E) Full length (fl) and soluble (s)PRR levels in the lysate (upper panel) and conditioned medium (lower panel) of furin knockout (furKO) and WT βTC3 cells. Three independent furKO clones were assessed. Quantifications on the right-hand side show n=3 independent experiments. *P

    Article Snippet: For deglycosylation experiments, 100 μg of proteins from the cell lysate were boiled at 95°C for 5 min in sodium phosphate buffer 0.1% SDS and 2-Mercaptoethanol (diluted 1:6 in water) in a total volume of 35ul.

    Techniques: Inhibition, Western Blot, Incubation, Concentration Assay, Transfection, Construct, Knock-Out, Clone Assay

    Comparison of changes in muscle protein synthesis and protein degradation between hindlimb muscles in response to unloading. Adult (9 mo) male rats underwent hindlimb unloading (HU) for 3, 7, or 14 days. Protein synthesis ( A ), 20S and 26S β 5 proteasome subunit activity ( B ), and cathepsin L activity ( C ) were measured in the soleus (open squares), medial gastrocnemius (MG; solid circles), and tibialis anterior (TA; shaded triangles) at each unloading time point. Total protein determined by stain-free imaging of the PVDF membrane was used to normalize puromycin protein expression. All values are expressed as a percentage relative to the control group for each muscle ( n = 5–6/muscle). Values are means ± SE. * P

    Journal: Journal of Applied Physiology

    Article Title: Muscle-specific and age-related changes in protein synthesis and protein degradation in response to hindlimb unloading in rats

    doi: 10.1152/japplphysiol.00703.2016

    Figure Lengend Snippet: Comparison of changes in muscle protein synthesis and protein degradation between hindlimb muscles in response to unloading. Adult (9 mo) male rats underwent hindlimb unloading (HU) for 3, 7, or 14 days. Protein synthesis ( A ), 20S and 26S β 5 proteasome subunit activity ( B ), and cathepsin L activity ( C ) were measured in the soleus (open squares), medial gastrocnemius (MG; solid circles), and tibialis anterior (TA; shaded triangles) at each unloading time point. Total protein determined by stain-free imaging of the PVDF membrane was used to normalize puromycin protein expression. All values are expressed as a percentage relative to the control group for each muscle ( n = 5–6/muscle). Values are means ± SE. * P

    Article Snippet: Cathepsin L enzymatic activity was assayed by adding 100 µM Z-Phe-Arg-AMC (Calbiochem, catalog no. 03321501) to a reaction buffer containing 100 mM sodium acetate, pH 5.5, 1 mM EDTA, and 2 mM DTT in the absence and presence of 10 μM cathepsin L inhibitor I (Calbiochem, catalog no. 219421).

    Techniques: Activity Assay, Staining, Imaging, Expressing

    Proteasome and cathepsin L activity changes in adult and old rats during hindlimb unloading (HU). Proteolytic activity of the β 5 -subunits of the 20S (ATP-independent) and 26S (ATP-dependent) proteasome and cathepsin L activity were measured in the soleus ( A ), medial gastrocnemius (MG; B ), and tibialis anterior (TA; C ) muscles of adult (9 mo, □) and old (29 mo, ■) rats after 3, 7, and 14 days of HU. Data are expressed as a percentage relative to the activity of the adult control group ( n = 4–6/group). Values are means ± SE. * P

    Journal: Journal of Applied Physiology

    Article Title: Muscle-specific and age-related changes in protein synthesis and protein degradation in response to hindlimb unloading in rats

    doi: 10.1152/japplphysiol.00703.2016

    Figure Lengend Snippet: Proteasome and cathepsin L activity changes in adult and old rats during hindlimb unloading (HU). Proteolytic activity of the β 5 -subunits of the 20S (ATP-independent) and 26S (ATP-dependent) proteasome and cathepsin L activity were measured in the soleus ( A ), medial gastrocnemius (MG; B ), and tibialis anterior (TA; C ) muscles of adult (9 mo, □) and old (29 mo, ■) rats after 3, 7, and 14 days of HU. Data are expressed as a percentage relative to the activity of the adult control group ( n = 4–6/group). Values are means ± SE. * P

    Article Snippet: Cathepsin L enzymatic activity was assayed by adding 100 µM Z-Phe-Arg-AMC (Calbiochem, catalog no. 03321501) to a reaction buffer containing 100 mM sodium acetate, pH 5.5, 1 mM EDTA, and 2 mM DTT in the absence and presence of 10 μM cathepsin L inhibitor I (Calbiochem, catalog no. 219421).

    Techniques: Activity Assay

    Cell cycle, apoptotic and proteolytic profiles in MDC1A and Surf1-LS muscle cells. ( A ) Cell distribution in the different phases of the cell cycle. ( B ) Differences in gene expression of CASP3, PSMA2, MURF1, MAFBX and USP19 in MDC1A and Surf1-LS myoblasts and myotubes compared to control. ( C ) Caspase-3 enzymatic activity was increased in myotubes from MDC1A and Surf1-LS patients compared to control cells. ( D ) 20S proteasome enzymatic activity was increased in both myoblasts and myotubes from MDC1A patients but was unchanged in Surf1-LS muscle cells compared to control. Results are expressed as a fold change of controls. Columns represent mean values and bars SE. Statistical significance was assessed by one-way ANOVA followed by Duncan’s post hoc test. p

    Journal: Scientific Reports

    Article Title: Bioenergetic Impairment in Congenital Muscular Dystrophy Type 1A and Leigh Syndrome Muscle Cells

    doi: 10.1038/srep45272

    Figure Lengend Snippet: Cell cycle, apoptotic and proteolytic profiles in MDC1A and Surf1-LS muscle cells. ( A ) Cell distribution in the different phases of the cell cycle. ( B ) Differences in gene expression of CASP3, PSMA2, MURF1, MAFBX and USP19 in MDC1A and Surf1-LS myoblasts and myotubes compared to control. ( C ) Caspase-3 enzymatic activity was increased in myotubes from MDC1A and Surf1-LS patients compared to control cells. ( D ) 20S proteasome enzymatic activity was increased in both myoblasts and myotubes from MDC1A patients but was unchanged in Surf1-LS muscle cells compared to control. Results are expressed as a fold change of controls. Columns represent mean values and bars SE. Statistical significance was assessed by one-way ANOVA followed by Duncan’s post hoc test. p

    Article Snippet: Caspase-3 enzymatic assay Caspase-3 activity was measured using the Caspase-3 Colorimetric Activity Assay Kit (Millipore).

    Techniques: Expressing, Activity Assay