Plasmids were constructed to encode the four different NAA10 (NM_003491.4) variants; NAA10 c.16G>C p.(A6P), NAA10 c.235C>T p.(R79C), NAA10 c.386A>C p.(Q129P), and NAA10 c.469G>A p.(E157K). The mutations were incorporated into a pcDNA3.1/NAA10-WT-V5 vector [previously called phARD1-V5 (Arnesen et al. 2005 (link))] using Q5 Site Directed Mutagenesis Kit (New England Biolabs). The primers used for mutagenesis are available in Supplementary Table S3. The resulting plasmids were verified by Sanger sequencing. Primary antibodies used in this study were V5-tag mouse monoclonal antibody (1:5000 dilution, Invitrogen R960-25), β-tubulin mouse monoclonal antibody (1:3000 dilution, Sigma-Aldrich T5293) and NAA15 rabbit polyclonal antibody (1:2000, Biogenes, previously called anti-NATH) (Arnesen et al. 2005 (link)). Secondary antibodies used were Mouse IgG HRP-Linked Whole Ab (1:5000 dilution, Cytiva NA931) and Rabbit IgG HRP-Linked Whole Ab (1:5000 dilution, Cytiva NA934).
Mouse igg hrp linked whole ab
Manufactured by Cytiva
The Mouse IgG HRP-Linked Whole Ab is a laboratory reagent used for the detection of mouse immunoglobulin G (IgG) in various immunoassays. It consists of a whole antibody molecule that is conjugated with horseradish peroxidase (HRP), an enzyme that can be used to generate a colorimetric or chemiluminescent signal upon the addition of a suitable substrate.
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Lab products found in correlation
Rabbit igg hrp linked whole ab, by Cytiva (2 mentions)
V5 tag mouse monoclonal antibody, by Thermo Fisher Scientific (1 mentions)
Q5 site directed mutagenesis kit, by New England Biolabs (1 mentions)
Ab26271, by Abcam (1 mentions)
Ab187650, by Abcam (1 mentions)
Ab113205, by Abcam (1 mentions)
Ab138022, by Abcam (1 mentions)
Pierce ecl plus western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Anti flag m2 antibody, by Merck Group (1 mentions)
2 protocols using mouse igg hrp linked whole ab
1
Construction and Validation of NAA10 Variants
2
Antibody-based Protein Analysis Techniques
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Antibodies used for protein analysis were as follows: Anti-FLAG M2 antibody (F3165 Sigma-Aldrich), Anti-ILF2 antibody (ab113205 Abcam), Anti-SAFB antibody [EPR13588] (ab187650 Abcam), Anti-RNA Helicase A antibody (ab26271 Abcam), Anti-HNRPA3/HNRNPA3 antibody (ab78300 Abcam), Anti-Matrin-3 antibody [EPR10635(B)] (ab151714 Abcam), Anti-SFRS7/SRSF7 antibody (ab138022 Abcam), hnRNP C1/C2 (D6S3N) Rabbit mAb (#91,327 Cell Signaling Technology), RBMX/hnRNP G (D7C2V Cell Signaling Technology) Rabbit mAb (#14,794 Cell Signaling Technology), PTBP1 (E4I3Q) Rabbit mAb (#57,246 Cell Signaling Technology), hnRNP K (R332) Antibody (#4675 Cell Signaling Technology), Anti-hnRNP A1 Antibody (4B10) (sc-32301 Santa Cruz Biotechnology), Anti-Histone H3 Antibody, CT, pan (07–690 Sigma-Aldrich), Anti-Lamin B1 antibody [EPR8985(B)] (ab133741 Abcam). The primary antibodies were detected using Rabbit IgG HRP Linked Whole Ab (NA934 Cytiva) or Mouse IgG HRP Linked Whole Ab (NA931 Cytiva). The bands were visualized using Pierce ECL Plus Western Blotting Substrate (Thermo Fisher Scientific).
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