Whole tissue lysates were prepared from ovaries and heads using RIPA buffer (Sigma Aldrich), and the protein concentration was measured using Pierce BCA Protein Assay Kit (ThermoFisher). Equal amounts of total protein extract were loaded as samples for western blot. Instead of using housekeeping genes as loading controls, we utilized the total protein normalization (TNP) method (Revert™700 Total Protein Stain Li-Cor). The TNP staining gives a linear signal range, which is especially relevant for low-abundance proteins such as Tj. Anti-DYKDDDDK-HRP antibodies (Miltenyi Biotec) were used to immunostain Tj-TR-3xFlag bands. HRP signal was developed using Chemiluminescent substrates (ThermoFisher) and imaged in the Odyssey Fc Imaging System. The anti-Flag chemiluminescent signal from each tissue derived from tjnat/nat mutants was normalized to the respective TNP signal and compared to the anti-Flag/TNP signal from tissues derived from tjTR/TR mutants to calculate TR efficiency of Tj.
Anti dykddddk hrp antibodies
Manufactured by Miltenyi Biotec
Anti-DYKDDDDK-HRP antibodies are a type of affinity reagent designed to detect and visualize proteins tagged with the DYKDDDDK (commonly known as 'FLAG') epitope. These antibodies are conjugated with horseradish peroxidase (HRP), which allows for sensitive and specific detection of the target proteins in various applications, such as Western blotting, immunoprecipitation, and enzyme-linked immunosorbent assays (ELISA).
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Lab products found in correlation
2 protocols using anti dykddddk hrp antibodies
1
Quantifying Tj Protein Levels in Drosophila
2
Quantitative Analysis of Tj Protein
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Whole tissue lysates were prepared from ovaries and heads using RIPA buffer (Sigma Aldrich), and the protein concentration was measured using Pierce TM BCA Protein Assay Kit. Equal amounts of total protein extract were loaded as samples for western blot. Instead of using housekeeping genes as loading controls, we utilized the total protein normalization (TNP) method (Licor 700 Total Protein Stain). The TNP staining gives a linear signal range, which is especially relevant for low-abundance proteins such as Tj. Anti-DYKDDDDK-HRP antibodies (Miltenyi Biotec) were used to immunostain Tj-TR-3xFlag bands. HRP signal was developed using Chemiluminescent substrates (ThermoFisher) and imaged in the Odyssey Fc Imaging System.
The anti-Flag chemiluminescent signal from each tissue derived from tj nat/nat mutants was normalized to the respective TNP signal and compared to the anti-Flag/TNP signal from tissues derived from tj TR/TR mutants to calculate TR efficiency of Tj.
The anti-Flag chemiluminescent signal from each tissue derived from tj nat/nat mutants was normalized to the respective TNP signal and compared to the anti-Flag/TNP signal from tissues derived from tj TR/TR mutants to calculate TR efficiency of Tj.
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