To detect the expression of Ki-67, the mRTEC cells were treated with CdCl2 (5 μM) with or without R-467 (1 μM), or S-467 (1 μM) for 24 h, or pre-treated with PD98059 (10 μM), SB202190 (10 μM) for 30 min. The cells were fixed for 30 min in 4% Formaldehyde (FA, Sigma-Aldrich). Then, the cells were permeabilized with 0.1% Triton X-100 (Sigma-Aldrich) in PBS for 20 min. After blocked with 3% normal goat serum, the cells were incubated with mouse anti-Ki-67 (1:100, Cell Signaling Technology) antibody overnight at 4 °C, followed by 1 h incubation with Alexa Fluor 488 goat anti-mouse IgG (1:200, Molecular Probes, Invitrogen). After washing twice with PBS, cell nucleus was stained by the DAPI (Invitrogen) for several minutes. The cells were washed in PBS for 10 min and the cells were mounted, then examined by Olympus IX73 microscopy (Japan). To detect the expression of CaSR (anti-CaSR, 1:40, Thermo fisher Scientific) on the membranes of HK-2 and mRTEC cells, the process was performed without the permeabilization step.
Anti casr
Manufactured by Thermo Fisher Scientific
The Anti-CaSR is a laboratory product that targets the Calcium-Sensing Receptor (CaSR) protein. It is designed for use in research applications involving the study of calcium homeostasis and related physiological processes.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Mouse anti ki67, by Cell Signaling Technology (1 mentions)
Formaldehyde fa, by Merck Group (1 mentions)
Ix73 microscopy, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti β tubulin, by Thermo Fisher Scientific (1 mentions)
Anti gr, by Thermo Fisher Scientific (1 mentions)
Multiskan go microplate spectrophotometer, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti casr
1
Ki-67 Expression in mRTEC Cells
2
Protein Expression Analysis in Rat Kidney and Thyroid
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Frozen kidney and total thyroid gland tissues obtained from all the rats were homogenized with RIPA lysis buffer in the presence of protease and phosphatase inhibitor cocktail. Protein concentrations were determined by BCA assay kit in Multiskan™ GO Microplate Spectrophotometer (Thermo Fisher Scientific; Paisley, England). Equal amounts of proteins were separated by sodium dodecyl sulfate polyacrylamide gels (SDS-PAGE) and were subsequently transferred to a PVDF membrane. Afterwards, the blots were incubated with the primary antibodies anti-PTH/PTHrP-R (PTHR1) (Santa Cruz), anti-CaSR (Thermo Scientific), anti-GR (Thermo Scientific), anti-β-tubulin (Thermo Scientific), and anti-β-actin (Santa Cruz). The blots were then exposed to chemiluminescence solution for visualization of the specific binding. Densitometric quantifications of the protein bands were done using ImageJ analysis system (NIH, Bethesda, USA). Results were normalized against the β-actin or β-tubulin expression in each group (15 (link)).
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