
| SKU | DZ41648 |
|---|---|
| Application | Western Blot |
| Sample | 3T3 Cell lysate |
| Sample Processing Description | Nuclear and Cytoplasmic Extraction was performed on 3T3 cells with NE-PER kit by following the protocol and by adding protease and phosphatase inhibitors. Total protein was quantified with Qubit Protein Assay. Lysates were mixed with 4× Laemmli Sample Buffer and 2-mercaptoethanol, and then heated at 95°C for 5 min. |
| Primary Antibody | Anti-Mouse Atxn7 Antibody |
| Primary Incubation | Incubated in non-fat dry milk in TBST overnight at 4°C with antibody dilution at 1:1000 dilution |
| Secondary Antibody | Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP |
| Secondary Incubation | 1:4000 dilution in milk for 1 hour at room temperature |
| Other Reagents used | SuperSignal West Pico PLUS Chemiluminescent Substrate |
| Detection | Biorad ChemiDoc |
| Results Summary | I was looking to target Atxn7 isoform 2, named as Atxn7b in the literature, and referenced as ENSMUST00000223714.2 on Ensembl. (It differs only on the C-terminus compared to Atxn7a, isoform 1). Atxn7 is mainly in the nucleus and is between 100-130 kDa. An shRNA targeting Atxn7 displayed a decrease in Atxn7 abondance showing specificity of the antibody. |










