The kidneys were isolated free from the surrounding connective tissue after sacrificing the mice. The kidney tissue was fixed with 4% paraformaldehyde, embedded in paraffin, and then cut into slices using a microtome (Leica RM 2235 or Leica CM1850UV; Leica, Solms, Germany). Immunohistochemistry was performed using a Histone Simple Stain Kit (Nichirei, Tokyo, Japan) according to the manufacturer's instructions. Briefly, serial sections (4 μm thick) were deparaffinized with xylene, and then gradient ethanol was used to dewax and hydrate the samples. The sections were immersed in methanol with 3% H2O2 for 15 min to inactivate endogenous peroxidases and then incubated with primary antibodies at room temperature for 1 h. Primary antibodies against NLRP3 (rabbit anti-NLRP3 antibody, 1 : 200; Proteintech) and caspase-1 (rabbit anti-caspase-1 anti-body, 1 : 200; Proteintech) were used. All sections were visualized using an Olympus microscope (Olympus, Tokyo, Japan).
Rabbit anti nlrp3 antibody
Manufactured by Proteintech
Sourced in China
The Rabbit anti-NLRP3 antibody is a laboratory reagent used for the detection and analysis of the NLRP3 protein. NLRP3 is a member of the NLR protein family and is involved in the regulation of the inflammatory response. This antibody can be used in various immunological techniques, such as Western blotting and immunohistochemistry, to study the expression and localization of the NLRP3 protein in biological samples.
Automatically generated - may contain errors
2 protocols using rabbit anti nlrp3 antibody
1
Immunohistochemical Analysis of NLRP3 and Caspase-1
2
NLRP3 Inflammasome Activation Pathway
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HDLECs were washed twice with phosphate-buffered saline and then lysed in RIPA buffer supplemented with PMSF (1 mM). Proteins were separated on SDS-PAGE gels and transferred onto the PVDF membrane (Immobilon, Massachusetts, USA). Membranes were blocked with 5% non-fat dry milk for 2 hr at room temperature and incubated with primary antibodies overnight at 4°C. Primary antibodies were as follows: rabbit anti-NLRP3 antibody (Proteintech, Wuhan, China, 1:1000), rabbit anti-caspase-1 antibody (Proteintech, 1:1000), rabbit anti-Casp1p20 antibody (Proteintech, 1:1000), mouse anti-Pro-IL-1β antibody (CST, Danvers, MA, USA; 1:1000) and mouse anti-IL-1β antibody (CST, 1:1000), Immunoreactivity was visualized with HRP-linked secondary antibodies (Millipore, Billerica, MA, USA) under a chemiluminometer.
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