This method was applied to the immuno-purified CalS complex. Proteins were denatured in LSB and separated by electrophoresis as described above; proteins were transferred to membranes and then underwent a process of denaturation/renaturation according to a protocol based on decreasing concentrations of guanidine-HCl (Yuliang et al. 2007 (link)). After overnight blocking, membranes were incubated with porcine tubulin (Cytoskeleton Inc.) at a concentration of 1 mg/mL in TBS supplemented with 0.1% Tween-20 (TBST) for 2 h at room temperature. After washing in TBS, membranes were incubated with primary anti-α-tubulin antibody B-5–1-2 (mouse) diluted 1:5000 in TBST or with antibody HDA (rabbit) against CalS diluted 1:300 in TBST. After a 1-h incubation at room temperature, membranes were washed in TBS and then incubated with secondary antibodies against mouse or rabbit immunoglobulins conjugated with HRP. Antibodies were used at a dilution of 1:5000 in TBST for 1 h at room temperature. After further washing in TBS, membranes were processed with the Clarity reagent (Bio-Rad) and the signal was detected with the Fluor-S device (Bio-Rad).
Fluor s device
Manufactured by Bio-Rad
The Fluor-S device is a versatile imaging system designed for fluorescence-based detection and analysis. It enables the visualization and quantification of fluorescently labeled biological samples, such as proteins, nucleic acids, and other molecules. The Fluor-S device utilizes a range of excitation and emission wavelengths to accommodate various fluorescent dyes and probes, allowing users to perform a wide variety of applications in the field of molecular biology and biochemistry.
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Lab products found in correlation
2 protocols using fluor s device
1
Immuno-purification and Western Blot Analysis
2
Immunopurified CaLS Complex Analysis
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This method was applied on the immuno-puri ed CalS complex. Proteins were denatured in Laemmli Sample Buffer (LSB) and separated by electrophoresis as described above; proteins were transferred to membranes and then underwent a process of denaturation/renaturation according to a protocol based on decreasing concentrations of guanidine-HCl (Yuliang et al. 2007 ). After overnight blocking, membranes were incubated with porcine tubulin (Cytoskeleton Inc.) at a concentration of 1 mg/mL in TBS supplemented with 0.1% Tween-20 (TBST) for 2 hours at room temperature. After washing in TBS, membranes were incubated with primary anti-α-tubulin antibody B-5-1-2 (mouse) diluted 1:5000 in TBST or with antibody HDA (rabbit) against CalS diluted 1:300 in TBST. After a one-hour incubation at room temperature, membranes were washed in TBS and then incubated with secondary antibodies against mouse or rabbit immunoglobulins conjugated with HRP. Antibodies were used at a dilution of 1:5000 in TBST for one hour at room temperature. After further washing in TBS, membranes were processed with the Clarity reagent (Bio-Rad) and the signal was detected with the Fluor-S device (Bio-Rad).
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