Enzyme-linked Immunosorbent Assay:Article Title: Cellular assay optimization: part I: the use of large-scale transiently transfected cryobanks and introduction of a c-Myc tag to design a standardized ELISA process.
Article Snippet: the 384-well tissue culture plates and 10-layer cellStacKS were supplied by corning incorporated (corning, ny), and the 384-well high-bind eliSa plates were supplied by greiner bio-one (monroe, nc). the 384-well poly-d-lysine-coated tissue culture plates for heK 293 cell plating were purchased from becton dickinson (bd) biosciences (franklin lakes, nJ). labcyte, inc. (Sunnyvale, ca) supplied the 384-well source plates for acoustic liquid dispensing. dulbecco’s modified eagle’s medium (dmem) and f12 medium were supplied by gibco invitrogen corp. (life Sciences limited, paisley, Scotland). fetal calf serum (fcS) and bovine serum albumin (bSa) were purchased from Sigma-aldrich company limited (dorset, england). opti-mem and lipofectamine 2000 reagents used for transfections were supplied by invitrogen, and pei, linear 25 kda (cat. #23966), was purchased from polysciences (Warrington, pa). lysis buffer comprised 25 mm tris/hcl, 3 mm edta, 3 m egta, 50 mm naf, 2 mm sodium orthovanadate, 0.27 m sucrose, 10 mm b-glycerophosphate, 5 mm pyrophosphate, and 0.5% (v/v) triton X-100. complete protease inhibitor tablets (roche, basel, Switzerland) were dissolved in lysis buffer 20 min before use. plate washer solution was prepared by adding 0.05% polysorbate and 10 ppm of proclin 300 (Sigmaaldrich) to phosphate-buffered saline (pbS)/a solution. faK primary antibody was purchased from bioSource (camarillo, ca), perK phospho–primary antibody was supplied by Santa cruz biotechnology (Santa cruz, ca), and dyrK phospho-antibody was manufactured in-house. antirabbit igg–horseradish peroxidase (hrp) linked antibody and the myc tag (9b11) capture antibody were purchased from cell Signaling technology (danvers, ma). bmX used streptavidinhrp supplied by ge healthcare (piscataway, nJ) and antiphosphotyrosine clone 4g-10-biotin conjugate from millipore (billerica, ma).
Modification:Article Title: Cellular assay optimization: part I: the use of large-scale transiently transfected cryobanks and introduction of a c-Myc tag to design a standardized ELISA process.
Article Snippet: the 384-well tissue culture plates and 10-layer cellStacKS were supplied by corning incorporated (corning, ny), and the 384-well high-bind eliSa plates were supplied by greiner bio-one (monroe, nc). the 384-well poly-d-lysine-coated tissue culture plates for heK 293 cell plating were purchased from becton dickinson (bd) biosciences (franklin lakes, nJ). labcyte, inc. (Sunnyvale, ca) supplied the 384-well source plates for acoustic liquid dispensing. dulbecco’s modified eagle’s medium (dmem) and f12 medium were supplied by gibco invitrogen corp. (life Sciences limited, paisley, Scotland). fetal calf serum (fcS) and bovine serum albumin (bSa) were purchased from Sigma-aldrich company limited (dorset, england). opti-mem and lipofectamine 2000 reagents used for transfections were supplied by invitrogen, and pei, linear 25 kda (cat. #23966), was purchased from polysciences (Warrington, pa). lysis buffer comprised 25 mm tris/hcl, 3 mm edta, 3 m egta, 50 mm naf, 2 mm sodium orthovanadate, 0.27 m sucrose, 10 mm b-glycerophosphate, 5 mm pyrophosphate, and 0.5% (v/v) triton X-100. complete protease inhibitor tablets (roche, basel, Switzerland) were dissolved in lysis buffer 20 min before use. plate washer solution was prepared by adding 0.05% polysorbate and 10 ppm of proclin 300 (Sigmaaldrich) to phosphate-buffered saline (pbS)/a solution. faK primary antibody was purchased from bioSource (camarillo, ca), perK phospho–primary antibody was supplied by Santa cruz biotechnology (Santa cruz, ca), and dyrK phospho-antibody was manufactured in-house. antirabbit igg–horseradish peroxidase (hrp) linked antibody and the myc tag (9b11) capture antibody were purchased from cell Signaling technology (danvers, ma). bmX used streptavidinhrp supplied by ge healthcare (piscataway, nJ) and antiphosphotyrosine clone 4g-10-biotin conjugate from millipore (billerica, ma).
Transfection:Article Title: Cellular assay optimization: part I: the use of large-scale transiently transfected cryobanks and introduction of a c-Myc tag to design a standardized ELISA process.
Article Snippet: the 384-well tissue culture plates and 10-layer cellStacKS were supplied by corning incorporated (corning, ny), and the 384-well high-bind eliSa plates were supplied by greiner bio-one (monroe, nc). the 384-well poly-d-lysine-coated tissue culture plates for heK 293 cell plating were purchased from becton dickinson (bd) biosciences (franklin lakes, nJ). labcyte, inc. (Sunnyvale, ca) supplied the 384-well source plates for acoustic liquid dispensing. dulbecco’s modified eagle’s medium (dmem) and f12 medium were supplied by gibco invitrogen corp. (life Sciences limited, paisley, Scotland). fetal calf serum (fcS) and bovine serum albumin (bSa) were purchased from Sigma-aldrich company limited (dorset, england). opti-mem and lipofectamine 2000 reagents used for transfections were supplied by invitrogen, and pei, linear 25 kda (cat. #23966), was purchased from polysciences (Warrington, pa). lysis buffer comprised 25 mm tris/hcl, 3 mm edta, 3 m egta, 50 mm naf, 2 mm sodium orthovanadate, 0.27 m sucrose, 10 mm b-glycerophosphate, 5 mm pyrophosphate, and 0.5% (v/v) triton X-100. complete protease inhibitor tablets (roche, basel, Switzerland) were dissolved in lysis buffer 20 min before use. plate washer solution was prepared by adding 0.05% polysorbate and 10 ppm of proclin 300 (Sigmaaldrich) to phosphate-buffered saline (pbS)/a solution. faK primary antibody was purchased from bioSource (camarillo, ca), perK phospho–primary antibody was supplied by Santa cruz biotechnology (Santa cruz, ca), and dyrK phospho-antibody was manufactured in-house. antirabbit igg–horseradish peroxidase (hrp) linked antibody and the myc tag (9b11) capture antibody were purchased from cell Signaling technology (danvers, ma). bmX used streptavidinhrp supplied by ge healthcare (piscataway, nJ) and antiphosphotyrosine clone 4g-10-biotin conjugate from millipore (billerica, ma).
Lysis:Article Title: Cellular assay optimization: part I: the use of large-scale transiently transfected cryobanks and introduction of a c-Myc tag to design a standardized ELISA process.
Article Snippet: the 384-well tissue culture plates and 10-layer cellStacKS were supplied by corning incorporated (corning, ny), and the 384-well high-bind eliSa plates were supplied by greiner bio-one (monroe, nc). the 384-well poly-d-lysine-coated tissue culture plates for heK 293 cell plating were purchased from becton dickinson (bd) biosciences (franklin lakes, nJ). labcyte, inc. (Sunnyvale, ca) supplied the 384-well source plates for acoustic liquid dispensing. dulbecco’s modified eagle’s medium (dmem) and f12 medium were supplied by gibco invitrogen corp. (life Sciences limited, paisley, Scotland). fetal calf serum (fcS) and bovine serum albumin (bSa) were purchased from Sigma-aldrich company limited (dorset, england). opti-mem and lipofectamine 2000 reagents used for transfections were supplied by invitrogen, and pei, linear 25 kda (cat. #23966), was purchased from polysciences (Warrington, pa). lysis buffer comprised 25 mm tris/hcl, 3 mm edta, 3 m egta, 50 mm naf, 2 mm sodium orthovanadate, 0.27 m sucrose, 10 mm b-glycerophosphate, 5 mm pyrophosphate, and 0.5% (v/v) triton X-100. complete protease inhibitor tablets (roche, basel, Switzerland) were dissolved in lysis buffer 20 min before use. plate washer solution was prepared by adding 0.05% polysorbate and 10 ppm of proclin 300 (Sigmaaldrich) to phosphate-buffered saline (pbS)/a solution. faK primary antibody was purchased from bioSource (camarillo, ca), perK phospho–primary antibody was supplied by Santa cruz biotechnology (Santa cruz, ca), and dyrK phospho-antibody was manufactured in-house. antirabbit igg–horseradish peroxidase (hrp) linked antibody and the myc tag (9b11) capture antibody were purchased from cell Signaling technology (danvers, ma). bmX used streptavidinhrp supplied by ge healthcare (piscataway, nJ) and antiphosphotyrosine clone 4g-10-biotin conjugate from millipore (billerica, ma).
Protease Inhibitor:Article Title: Cellular assay optimization: part I: the use of large-scale transiently transfected cryobanks and introduction of a c-Myc tag to design a standardized ELISA process.
Article Snippet: the 384-well tissue culture plates and 10-layer cellStacKS were supplied by corning incorporated (corning, ny), and the 384-well high-bind eliSa plates were supplied by greiner bio-one (monroe, nc). the 384-well poly-d-lysine-coated tissue culture plates for heK 293 cell plating were purchased from becton dickinson (bd) biosciences (franklin lakes, nJ). labcyte, inc. (Sunnyvale, ca) supplied the 384-well source plates for acoustic liquid dispensing. dulbecco’s modified eagle’s medium (dmem) and f12 medium were supplied by gibco invitrogen corp. (life Sciences limited, paisley, Scotland). fetal calf serum (fcS) and bovine serum albumin (bSa) were purchased from Sigma-aldrich company limited (dorset, england). opti-mem and lipofectamine 2000 reagents used for transfections were supplied by invitrogen, and pei, linear 25 kda (cat. #23966), was purchased from polysciences (Warrington, pa). lysis buffer comprised 25 mm tris/hcl, 3 mm edta, 3 m egta, 50 mm naf, 2 mm sodium orthovanadate, 0.27 m sucrose, 10 mm b-glycerophosphate, 5 mm pyrophosphate, and 0.5% (v/v) triton X-100. complete protease inhibitor tablets (roche, basel, Switzerland) were dissolved in lysis buffer 20 min before use. plate washer solution was prepared by adding 0.05% polysorbate and 10 ppm of proclin 300 (Sigmaaldrich) to phosphate-buffered saline (pbS)/a solution. faK primary antibody was purchased from bioSource (camarillo, ca), perK phospho–primary antibody was supplied by Santa cruz biotechnology (Santa cruz, ca), and dyrK phospho-antibody was manufactured in-house. antirabbit igg–horseradish peroxidase (hrp) linked antibody and the myc tag (9b11) capture antibody were purchased from cell Signaling technology (danvers, ma). bmX used streptavidinhrp supplied by ge healthcare (piscataway, nJ) and antiphosphotyrosine clone 4g-10-biotin conjugate from millipore (billerica, ma).
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