Pro–IL-1β (12703), cleaved IL-1β (83186), anti–gasdermin D (96458), cleaved gasdermin D (36425), and ASC (13833) rabbit monoclonal antibodies were purchased from Cell Signaling Technology. Rabbit anti-AIM2 (ab93015), cleaved IL-1β (ab2105), and NLRP3 (ab214185) were purchased from Abcam. Rabbit anti-GAPDH (sc-25778) and mouse anti–IL-1R (sc-393998) were obtained from Santa Cruz Biotechnology. Antirabbit HRP secondary antibody was purchased from Thermo Fisher Scientific (31460).
Nlrp3 ab214185
Manufactured by Abcam
Sourced in United Kingdom, United States
NLRP3 (ab214185) is a recombinant protein that corresponds to a portion of the human NLRP3 protein. NLRP3 is a member of the NLR (nucleotide-binding domain, leucine-rich repeat) family of proteins and is involved in the formation of the NLRP3 inflammasome, a multiprotein complex that plays a role in the immune response.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti gapdh sc 25778, by Santa Cruz Biotechnology (1 mentions)
Cleaved caspase 1, by Cell Signaling Technology (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Cleaved gsdmd, by Cell Signaling Technology (1 mentions)
Interleukin 6 (il 6), by Cell Signaling Technology (1 mentions)
Phospho iκbα, by Cell Signaling Technology (1 mentions)
Caspase 1, by Cell Signaling Technology (1 mentions)
Tnf α, by Cell Signaling Technology (1 mentions)
Il 1β, by Cell Signaling Technology (1 mentions)
Nf κb p65, by Cell Signaling Technology (1 mentions)
Phospho nf κb p65, by Cell Signaling Technology (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Caspase 1 2225, by Cell Signaling Technology (1 mentions)
Tlr4 ab13556, by Abcam (1 mentions)
Gapdh 2118, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3 9661, by Cell Signaling Technology (1 mentions)
Sodium orthovanadate, by Merck Group (1 mentions)
Sodium fluoride, by Merck Group (1 mentions)
3 protocols using nlrp3 ab214185
1
Detecting Inflammasome Activation Markers
2
Western Blot Analysis of Inflammatory Proteins
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Cells were lysed and total proteins were subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), transferred to a polyvinylidene difluoride membrane (Millipore, Germany). The membrane was sequential incubated with primary antibody and secondary antibody (LI-COR Co., USA), then detected by ODYSSEY infrared imaging system (LI-COR Co. USA). The primary antibodies including caspase-1 (#24232), cleaved-caspase-1(#89332), IL-1β (#12242), NF-κB P65 (#8242), phospho-NF-κB P65 (#3033), IκBα (#4814), phospho-IκBα (#2859), cleaved-GSDMD (#50928), TNF-α (#11948) and IL-6 (#12912) were purchased from Cell Signaling Technology, USA; GSDMD (ab239377), IL-18 (ab71495) and NLRP3 (ab214185) antibodies were purchased from Abcam, UK.
3
RAW264.7 Cell Protein Extraction and Western Blot
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Protein was extracted from RAW264.7 cells with RIPA buffer, containing 50 mM Tris-HCl (pH 7.4), 150 mM sodium chloride, 1% NP-40, 0.25% sodium deoxycholate, 1 mM EDTA, 1% protease inhibitors cocktail (78425, Pierce, Rockford, IL, USA), 1 mM PMSF (36978, Pierce), 1 mM sodium orthovanadate (S6508, Sigma Aldrich) and 50 mM sodium fluoride (S7920, Sigma Aldrich). Protein concentrations were determined using the BCA Protein Assay Kit (23227, Pierce). 40 μg of total protein was resolved by SDS-PAGE. The blots were probed with primary antibodies against (TLR4 (#ab13556) (1:2000), NLRP3 (#ab214185) (1:1000), (both from Abcam, Cambridge, MA, USA), ASC (#67824) (1:500), p-p65 (#3033) (1:1000), totalp65 (#8242) (1:1000), caspase-1 (#2225) (1:1000), pro-caspase-1 (#12242) (1:1000), cleaved caspase-3 (#9661) (1:1000) or GAPDH (#2118) (1:10000) (all from Cell Signaling Technology, Beverly, MA, USA).
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