Table 2. Incubation conditions and dilution factors of the different antibodies used in the present study.
| Denaturing conditions | Primary antibody dilution factor and blocking buffer* | Secondary antibody dilution factor† | |
|---|---|---|---|
| PEPT11 | 95°C, 5 min | 1:500; 5% BSA/TBST | 1:2,0008 |
| SGLT12 | 40°C, 15 min | 1:2,000; 5% M TBST | 1:20,0007 |
| pSGLT1 Ser418 3 | 40°C, 15 min | 1:200; 5% M TBST | 1:15,0007 |
| CFTR4 | 95°C, 5 min | 1:1,000; 5% M TBST | 1:2,0006 |
| Villin5 | 95°C, 5 min | 1:5,000; 5% TBST | 1:25,0009 |
* Incubated at 4°C overnight;
† Incubated at room temperature for 90 min.
1 monoclonal mouse-anti-PEPT1 (373742, Santa Cruz, lot no.: 31916).
2 polyclonal rabbit- anti-SGLT1 (ab14686, Abcam, UK, lot no.: JBC17887303).
3 monoclonal rabbit-antipSGLT1, custom made by Perbio Science, Germany (epitope: KIRKRApSEKELMI).
4 monoclonal rabbit-anti-CFTR (#78335, Cell Signaling Technology, lot no.: 1).
5 monoclonal mouse-anti-Villin (1D2C3, Santa Cruz, lot no.: L0808).
6 goat-anti-rabbit IgG HRP-conjugated (#7074, Cell Signaling Technology, lot no.: 26).
7 goat-anti-rabbit IgG HRP-conjugated (A9169, Sigma-Aldrich).
8 mouse-IgGκ BP-HRP-conjugated (516102, Santa Cruz).
9 goat-anti-mouse IgG HRP-conjugated (A2304, Sigma-Aldrich).
BSA: Bovine serum albumin; TBST: Tris-buffered saline including 0.1% Tween 20; M: Skimmed dry milk.