Cells or tissues were fixed in 4% paraformaldehyde (in 1× PBS), followed by permeabilization in 0.25% Triton X-100 for 15–30 min, then they were washed in PBS and incubated with primary antibody for at least 1 h at room temperature. Following washing in PBS, cells/tissues were incubated with Alexa Fluor–conjugated secondary antibodies. Alexa Fluor–conjugated phalloidin was purchased from Invitrogen (A12381). Rabbit anti–IL-1β (ab9722) and anti–caspase-1 (ab62698) antibodies were purchased from Abcam. Rabbit anti–gasdermin D (96458), cleaved gasdermin D (36425), and cleaved IL-1β (83186) were purchased from Cell Signaling Technology. Cells or tissues were mounted with VECTASHIELD mounting medium (H-1200; Vector Laboratories) containing DAPI, and images were captured using an Olympus IX83 inverted fluorescence microscope or a Zeiss LSM confocal microscope. Images were adjusted for brightness/contrast using Adobe Photoshop (Adobe). For three-dimensional sections, confocal images were acquired at 0.35–0.5-µm intervals.
Cleaved il 1β 83186
Manufactured by Cell Signaling Technology
Sourced in United States
Cleaved IL-1β (83186) is a laboratory product offered by Cell Signaling Technology. It is an antibody that detects the cleaved form of interleukin-1 beta (IL-1β), a pro-inflammatory cytokine. The antibody can be used to identify and quantify the cleaved form of IL-1β in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor conjugated phalloidin, by Thermo Fisher Scientific (1 mentions)
Ab62698, by Abcam (1 mentions)
Rabbit anti il 1β ab9722, by Abcam (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
Caspase 3 9662, by Cell Signaling Technology (1 mentions)
Parp 9542, by Cell Signaling Technology (1 mentions)
Hif 1α 36169, by Cell Signaling Technology (1 mentions)
Cy3 labeled goat anti rabbit igg h l a0516, by Beyotime (1 mentions)
Ab180799, by Abcam (1 mentions)
Nlrp3 15101, by Cell Signaling Technology (1 mentions)
Cy3 labeled goat anti mouse igg h l a0521, by Beyotime (1 mentions)
Hrp conjugated affinipure goat anti rabbit igg sa00001 2, by Proteintech (1 mentions)
Lc3ii 14600 1 ap, by Proteintech (1 mentions)
Gapdh 60004 1 ig, by Proteintech (1 mentions)
Ab93015, by Abcam (1 mentions)
3 protocols using cleaved il 1β 83186
1
Fluorescent Immunolabeling of Inflammatory Proteins
2
Western Blot Analysis of Inflammatory Markers
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After extraction, proteins in cell lysates were first resolved by SDS–PAGE, and then transferred to nitrocellulose membrane and subsequently incubated with the primary antibody. The antibodies to HIF-1α (36169; 1:1000), pro-IL-1β (12242; 1:1000), Cleaved IL-1β (83186; 1:1000), Caspase-3 (9662; 1:1000), PARP (9542s; 1:1000) were obtained from Cell Signaling Technology. The antibody to Caspase-1 (AG-20B-0042, 1:1000) was obtained from Adipogen. The antibody to NLRP3(IMG-6668A; 1:1000) was obtained from Novus. The antibody to α-Tubulin (11224-1-AP, 1:3000) was obtained from Proteintech. After incubation with Horseradish peroxidase-conjugated secondary antibodies, the signals were visualized by enhanced chemiluminescence (Pierce, Rockford, IL, USA, 32106) according to the manufacturer's instruction. The relative band intensity was quantified using the ImageJ Analyzer software (Media Cybernetics, Bethesda, MD, USA).
3
Antibody Sources for Cellular Assays
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Antibodies against CDKN1A (10355-1-AP), interleukin (IL)-18 (10663-1-AP), cleaved poly (ADP-ribose) polymerase (C-PARP, 13371-1-AP), glutathione peroxidase 4 (GPX4, 67763-Ig), HMGB1 (10829-1-AP), lipidation of microtubule-associated protein 1 light chain 3 form II (LC3-II, 14600-1-AP), P53BP1 (20002-1-AP), and GAPDH (60004-1-Ig) were obtained from Proteintech (Wuhan, China). Antibodies against caspase-3 (CASP-3, D3R6Y), cleaved-IL-1β (83186), γH2AX (9718), nucleotide-binding and oligomerization domain-like receptor (NLR) pyrin domain-containing 3 (NLRP3, 15101), and cleaved-caspase-1 (89332) were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies against absent in melanoma 2 (AIM2, ab93015), gasdermin-D (GSDMD)-N (ab215203), and apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC, ab180799) were purchased from Abcam (Massachusetts, UK). Horseradish peroxidase (HRP)-conjugated AffiniPure goat anti-mouse IgG (SA00001-1) and HRP-conjugated AffiniPure goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech. Cy3-labeled goat anti-mouse IgG (H + L) (A0521) and Cy3-labeled goat anti-rabbit IgG (H + L) (A0516) were purchased from Beyotime (Shanghai, China).
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