Mouse tissues were lysed using a protein lysis buffer containing 20 mM Tris (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100, 25 mM sodium pyrophosphate, and 2 mM sodium orthovanadate aprotinin. All denatured proteins were separated on an SDS-PAGE gel and then transferred to polyvinylidene difluoride membranes (Roche, Netley, NJ, United States). The membranes were blocked with 5% skimmed milk in Tris-buffered saline and then were incubated with 1:500 dilutions of primary antibodies as follows: anti-FGA (Abcam, Cambridge, MA, United States), anti-Slc12a1 (Abcam, Cambridge, MA, United States), and anti-Havcr1 (Abcam, Cambridge, MA, United States). Then, the samples were incubated with a horseradish peroxidase-conjugated anti-rabbit secondary antibody (Jackson ImmunoResearch, PA, United States). The bands were visualized using an ECL Western Blotting Kit (Biovision, Milpitas, CA, United States) and were quantified by Quantity One software (Bio-Rad, Hercules, CA, United States).
Anti slc12a1
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-Slc12a1 is a primary antibody that targets the Slc12a1 protein. Slc12a1 is a sodium-potassium-chloride cotransporter involved in ion and water homeostasis. The antibody can be used to detect and study the expression of Slc12a1 in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti fga, by Abcam (1 mentions)
Ecl western blotting kit, by Abcam (1 mentions)
Anti rabbit horseradish peroxidase conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
Polyvinylidene difluoride membrane, by Roche (1 mentions)
Dako antibody diluent, by Agilent Technologies (1 mentions)
Dab chromogen kit, by Agilent Technologies (1 mentions)
2 protocols using anti slc12a1
1
Western Blot Analysis of Mouse Tissue Proteins
2
Immunohistochemical Quantification of Thick Ascending Limb
Check if the same lab product or an alternative is used in the 5 most similar protocols
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A formalin-fixed and paraffin-embedded 3-μm-thick kidney specimens were placed on ultra plus glass slides. After rehydration, the target antigen epitope heat retrieval was performed on the sections at pH 6 and 95°C for 20 minutes. After blocking endogenous peroxidase, the antigen was detected with the rabbit anti-SLC12A1 (2.5 μg/mL; Abcam, Cambridge, United Kingdom, Cat. #ab191315, dilution 1:100) at 4°C overnight. The immunoreactivity was detected with a secondary antibody (Flex+Mouse Linker) for 30 minutes at room temperature. Both the first and the second antibodies were diluted with the DAKO antibody diluent containing background-reducing components (DAKO, Carpinteria, CA, USA). A DAB chromogen kit (DAKO, Glostrup, Denmark) was used for visualization of immunoreactivity. The sections were counterstained with Mayer hematoxylin, cleared, mounted, and studied under the microscope. The negative control was performed without applying the primary antibody in 1 slide in each of the series of stained slides. Four high-power images of the outer medulla were obtained to measure the height of epithelial lining cells (n = 50 per kidney) of a thick ascending limb (TAL) of the loop of Henle.
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