The premade blots are prepared with the appropriate GenLysate and 50μg protein is loaded onto a 4-20% denaturing polyacrylamide gel, along with a prestained molecular weight marker. After the proteins are resolved they are transferred to a PVDF membrane, using Efficient Western Transfer Buffer.
The blots are ready to be blocked and probed with the antibodies of choice.
Note: We recommend that customers contact our technical department for the latest information on the blots as G-Biosciences reserves the right to change the blot profile due to the availability of GenLysate.
Each blot has multiple tissue types from either human, mouse or rat species.
Multi tissue blots permit researchers to visualize the tissue distribution of their protein in a particular species. These have been successfully used for the analysis of a wide variety of proteins. For example, JIP3, a scaffold protein of the JNK pathway, had a specific brain location and was confirmed to be a neuronal protein (1); human Cds1-related kinase had a testicular localization and was shown to be a meiotic checkpoint kinase (2); the TATA-box binding protein related factor was present in all human tissues (3); Geminin, found solely in testis, was shown to be localized to proliferating cells (6).
An example of our multi tissue; single species blots is shown below (Figure 1). The mouse multi tissue blot (Cat. # TB38) was probed with antibodies against caveolin and the human multi tissue blot (Cat. # TB37-I) was probed with Cox-2.
Blot lane order:
- Protein marker
- Human Normal Uterus Lysate
- Human Normal Breast Lysate
- Human Normal Cervix Lysate
- Human Normal Rectum Lysate
- Human Normal Prostate Lysate
- Human Normal Thyroid Lysate
- Human Normal Hypopharynx Lysate
- Human Normal Stomach Lysate
- Human Normal Skin Lysate