goat anti digoxigenin antibody  (Vector Laboratories)

Journal: PLoS ONE

Article Title: A Simple, Sensitive and Safe Method to Determine the Human α/β-Tryptase Genotype

doi: 10.1371/journal.pone.0114944

Figure Lengend Snippet: Genomic DNA extracts from HMC-1, Mac-6 and two SMC preparations were each tested for the presence of β and α-tryptase genes by three different methods, each using the same 5′ and 3′ PCR primer pair as described in . When the 1017-28 bp amplimers were exposed to EcoRV, the α-tryptase amplimer yielded 678 and 339-40 bp fragments. After labeling with ethidium bromide, all such products were detected (upper panel). In contrast, only the high molecular weight amplimer and 339-40 bp fragment were visualized using either DY682- (middle panels) or digoxigenin- (lower panels) labeled 3′-primer. DNA sequence analyses of the restriction site are shown above the corresponding gel electrophoresis pattern.

Article Snippet: Betaine (B0300), Me 2 SO (DMSO), glycerin, phosphate-buffered saline, 2-( N -morpholino)ethane sulfonic acid (MES), Hepes, Tris, EDTA, bovine serum albumin, agarose, and MgCl 2 (Sigma-Aldrich Chemical Company LLC, St. Louis, MO); dNTP mix, gDNA purification kit, RNase-free DNase, and EcoRV (Promega Company, Fitchburg, WI); ProbeQuant G-50 Micro column (Amersham, Piscataway, NJ); FastStart Taq DNA Polymerase (Roche Diagnostics, Indianapolis, IN); polyacrylamide gels (Invitrogen, Carlsbad, CA); goat affinity-purified polyclonal IgG anti-digoxigenin (Vector Laboratories, Burlingame, CA); and IRDye680RD-labeled affinity-purified donkey IgG anti-goat IgG (LiCor, Lincoln, NE) were obtained as indicated.

Techniques: Labeling, Molecular Weight, Sequencing, Nucleic Acid Electrophoresis

Journal: PLoS ONE

Article Title: A Simple, Sensitive and Safe Method to Determine the Human α/β-Tryptase Genotype

doi: 10.1371/journal.pone.0114944

Figure Lengend Snippet: Purified gDNA from a SMC preparation having both α- and β- tryptase genes and from MAC-6 cells, in each case at doses ranging from 0.02 to 81.9 ng, were subjected to PCR and labeled using ethidium bromide (top panels), or to PCR with DY682- (middle panels) or digoxigenin- (bottom panels) labeled 3′-primer. In each case labeled 1017-28 bp amplimer bands are shown along with the net fluorescence in the corresponding plots, representing three independent experiments (data in ).

Article Snippet: Betaine (B0300), Me 2 SO (DMSO), glycerin, phosphate-buffered saline, 2-( N -morpholino)ethane sulfonic acid (MES), Hepes, Tris, EDTA, bovine serum albumin, agarose, and MgCl 2 (Sigma-Aldrich Chemical Company LLC, St. Louis, MO); dNTP mix, gDNA purification kit, RNase-free DNase, and EcoRV (Promega Company, Fitchburg, WI); ProbeQuant G-50 Micro column (Amersham, Piscataway, NJ); FastStart Taq DNA Polymerase (Roche Diagnostics, Indianapolis, IN); polyacrylamide gels (Invitrogen, Carlsbad, CA); goat affinity-purified polyclonal IgG anti-digoxigenin (Vector Laboratories, Burlingame, CA); and IRDye680RD-labeled affinity-purified donkey IgG anti-goat IgG (LiCor, Lincoln, NE) were obtained as indicated.

Techniques: Purification, Labeling, Fluorescence

Journal: PLoS ONE

Article Title: A Simple, Sensitive and Safe Method to Determine the Human α/β-Tryptase Genotype

doi: 10.1371/journal.pone.0114944

Figure Lengend Snippet: EcoRV-digested amplimers were labeled with ethidium bromide after gel electrophoresis (top panels) or, during the final PCR cycle with DY682 (middle panels) or digoxigenin (bottom panels) labeled 3′-primer. gDNA from SMCs with known ββββ, βββα and ββαα genotypes, defined as 4∶0, 3∶1 and 2∶2 β:α ratios, were utilized. Experimentally calculated percentages of the β-tryptase gene were plotted against the known percentages of β-tryptase (n = 3, mean ± SD). Standard deviations were too small to visualize the error bars in the 50 and 75%β-Tryptase (theoretical) values for the middle plot. Data is stored in .

Article Snippet: Betaine (B0300), Me 2 SO (DMSO), glycerin, phosphate-buffered saline, 2-( N -morpholino)ethane sulfonic acid (MES), Hepes, Tris, EDTA, bovine serum albumin, agarose, and MgCl 2 (Sigma-Aldrich Chemical Company LLC, St. Louis, MO); dNTP mix, gDNA purification kit, RNase-free DNase, and EcoRV (Promega Company, Fitchburg, WI); ProbeQuant G-50 Micro column (Amersham, Piscataway, NJ); FastStart Taq DNA Polymerase (Roche Diagnostics, Indianapolis, IN); polyacrylamide gels (Invitrogen, Carlsbad, CA); goat affinity-purified polyclonal IgG anti-digoxigenin (Vector Laboratories, Burlingame, CA); and IRDye680RD-labeled affinity-purified donkey IgG anti-goat IgG (LiCor, Lincoln, NE) were obtained as indicated.

Techniques: Labeling, Nucleic Acid Electrophoresis