Mil 1β

Manufactured by Abcam

MIl-1β is a recombinant human interleukin-1 beta protein. Interleukin-1 beta is a pro-inflammatory cytokine involved in the regulation of immune and inflammatory responses.

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Lab products found in correlation

Citrullinated histone h3, by Abcam (2 mentions) Mcaspase 1, by Adipogen (2 mentions) Mnlrp6, by Merck Group (2 mentions) Hnlrp6, by Abcepta (2 mentions) Hnlrp3, by Cell Signaling Technology (2 mentions) Complete protease phosphatase inhibitor, by Roche (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Triton x 100, by Roche (2 mentions) Normal goat serum (ngs), by Beyotime (1 mentions) Nlrp3, by Beyotime (1 mentions) Image pro plus 6, by Media Cybernetics (1 mentions) Caspase 1, by Abcam (1 mentions) 3 3 diaminobenzidine dab, by ZSGB-BIO (1 mentions) Cleaved caspase 1, by Cell Signaling Technology (1 mentions) Pro caspase 1, by Abcam (1 mentions) Pro il 1β, by Abcam (1 mentions) Gapdh, by ABclonal (1 mentions)

4 protocols using mil 1β

Immunohistochemical Evaluation of NLRP3, Caspase-1, and IL-1β

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Sections were permeabilized with 1% Triton X-100 for 2 h and blocked with normal goat serum (Beyotime Institute of Biotechnology, Haiman, China) for 30 min at room temperature, the sections were incubated sequentially with NLRP3 (1:500; cat. no. ab223687; Abcam), caspase-1 (1:500; cat. no. ab108362; Abcam) and mIL-1β (1 µg/ml; cat. no. ab9722; Abcam) antibodies at 4°C overnight. The next day, after rewarming for 1 h, sections were washed with PBS and then incubated with mouse anti-rabbit IgG-HRP (cat. no. sc-2357, 1:100) antibody for 2 h at room temperature. To visualize the signals, sections were treated with peroxidase substrate 3,3′-diaminobenzidine (DAB, 0.05%, ZSGB-Bio, China) and counterstained with hematoxylin for 1 min at room temperature. Sections were viewed and imaged under a light microscope (Ni-U; Nikon Corporation, Tokyo, Japan). Images were analyzed quantitatively using Image-Pro Plus 6.0 (Media Cybernetics, Inc., Rockville, MD, USA).

Qiao Y., Tian X., Men L., Li S., Chen Y., Xue M., Hu Y., Zhou P., Long G., Shi Y., Liu R., Liu Y., Qi Z., Cui Y, & Shen Y. (2018). Spleen tyrosine kinase promotes NLR family pyrin domain containing 3 inflammasome-mediated IL-1β secretion via c-Jun N-terminal kinase activation and cell apoptosis during diabetic nephropathy. Molecular Medicine Reports, 18(2), 1995-2008.

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Molecular Mechanisms of NLRP3 Inflammasome Activation

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Briefly, the proteins from H9c2 cells or heart tissues were separated by 12% SDS‐polyacrylamide gels and transferred onto PVDF membranes. After blocked with TBST containing 5% BSA, membranes were incubated with NLRP3, ASC, TLR6 (Santa Cruz), Pro‐IL‐1β, Pro‐Caspase 1 (abcam), mIL‐1β, Cleaved Caspase‐1, p65, p‐p65, IKKα, IKKβ, p‐IKKα/β, p‐TAK1, TAK1, p‐JNK, JNK, p‐MKK3/6, MKK6, p‐p38, p38, VDAC (Cell Signaling Technology Inc), Cytochrome C (Bioword), UCP2 (Proteintech) rabbit antibodies and GAPDH (ABclonal Technology) mouse antibody overnight at 4℃. Then, the membranes were incubated with horseradish peroxidase‐labelled secondary antibodies (ABclonal Technology) for 2 hours at room temperature. Subsequently, the protein bands were detected with Pierce™ ECL Western Blotting Substrate and scanned by an automatic chemiluminescence imaging system (Tanon 5200; Tanon).

Zhang H., Ma Y., Cao R., Wang G., Li S., Cao Y., Zhang H., Liu M., Liu G., Zhang J., Li S., Wang Y, & Ma Y. (2020). Soluble uric acid induces myocardial damage through activating the NLRP3 inflammasome. Journal of Cellular and Molecular Medicine, 24(15), 8849-8861.

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Lung Cytokine Profiling by ELISA and Western Blot

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Western blotting for lung homogenates and cell lysates was performed as described in earlier publications^{22 , 30 (link)}. Antibodies to mCaspase-1 (Adipogen), mIl-1β (Abcam), mNLRP6 (Sigma), hNLRP6 (Abgent), hNLRP3 (Cell Signaling), citrullinated histone H3 (Abcam) and GAPDH (Cell signaling) were used for immunoblotting. Human IL-1β and IL-18 and mouse TNF-α, MCP-1, IL-1β, IL-6, G-CSF, and IL-17A (eBioscience), and mouse CXCL1, CXCL2, and CXCL5 (R&D systems) were quantified according to the manufacturers’ protocols. To perform ELISA in mouse lung homogenates, snap frozen lung lobes were thawed, homogenized in 2 mL PBS containing 0.1% triton X-100 supplemented with complete protease/phosphatase inhibitor (1 tablet/50 mL) (Roche), centrifuged, and filtered prior to use for ELISA. Readings were normalized to total protein of lung homogenates and expressed as pg/mg.

Cai S., Paudel S., Jin L., Ghimire L., Taylor C.M., Wakamatsu N., Bhattarai D, & Jeyaseelan S. (2020). NLRP6 Modulates Neutrophil Homeostasis in Bacterial Pneumonia-Derived Sepsis. Mucosal immunology, 14(3), 574-584.

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Lung Cytokine Profiling by ELISA and Western Blot

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