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.
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.
- Protein marker
- Human Normal Liver Lysate
- Human Normal Brain Lysate
- Human Normal Lung Lysate
- Human Normal Kidney Lysate
- Human Normal Spleen Lysate
- Human Normal Testis Lysate
- Human Normal Ovary Lysate
- Human Normal Heart Lysate
- Human Normal Pancreas Lysate
- Hayashi, H. et al (2012) J Biol Chem 2012; 287:15054
- Tan, W. et al (2005) Nuc. Acid Res. 33: 3855
- Tohyama, K. et al (2004) Endocrinology. 145: 4384-4391
- Persengiev, S.P. et al (2003) PNAS. 100:14887
- Tuynder, M., et al (2002) PNAS. 99:14976
- Bernard, D. et al (2002) RNA 8: 526
- Wohlschlegel, J. A., et al (2002) Am. J. Pathol. 161: 267
- Kelkar, N. et al (2000) Mol. Cell. Biol. 20:1030
- Boettner, B. et al (2000) PNAS. 97: 9064
- Brown, A. L. et al (1999) PNAS. 96:3745