AA (purity > 98%) from Chengdu Purechem-Standard Co., Ltd. (Chengdu, China) was dissolved in 0.5% sodium carboxymethyl cellulose (CMC-Na) solution. Dexamethasone acetate (Dex) was from Tianjin Tianyao Pharmaceuticals (Tianjin, China) and dissolved in normal saline with 1% DMSO. Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The myeloperoxidase (MPO) assay kit was obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). TNF-α, IL-6, and IL-1β ELISA kits were purchased from BioLegend Inc. (CA, USA). All antibodies used in this study were supplied by Cell Signaling Technology Inc. (Beverly, MA).
Il 1β elisa kit
Manufactured by BioLegend
Sourced in United States
The IL-1β ELISA kits are designed for the quantitative measurement of IL-1β in biological samples. They utilize the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify the target analyte.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 6 (il 6), by BioLegend (2 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Myeloperoxidase mpo assay kit, by Nanjing Jiancheng (1 mentions)
Antibodies used in this study, by Cell Signaling Technology (1 mentions)
Rvd1 elisa kit, by Cayman Chemical (1 mentions)
Elisa plate reader, by Bio-Rad (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Tnf α, by BioLegend (1 mentions)
Lps escherichia coli o55 b5, by Merck Group (1 mentions)
Minicomplete protease inhibitors, by Roche (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Tnf α and il 6 kits, by R&D Systems (1 mentions)
Infinite m1000 pro, by Tecan (1 mentions)
Il 8 elisa kit, by BD (1 mentions)
Masson s trichrome stain kit, by Solarbio (1 mentions)
Alexa fluor 488 secondary antibody, by Abcam (1 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions)
Anti ocn antibody, by ABclonal (1 mentions)
Anti il 1β, by Abcam (1 mentions)
10 protocols using il 1β elisa kit
1
Anti-inflammatory Agent Evaluation
2
Quantification of IL-1β and RvD1 in Serum and CSF
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IL-1β ELISA kits were from BioLegend (Catalog number, 432604) and the assay was conducted according to the manufacturer’s protocol. RvD1 ELISA kits from Cayman Chemicals (Catalog number, 500380) were used. For each reaction, 50 μl of serum (collected from tail vein) and 20 μl of CSF (collected from cisterna magna) were collected. Each ELISA was conducted according to the manufacturer’s instruction, and standard curves were included each time. RvD1 ELISA was conducted as previously described (Zhang et al., 2018 (link); Tao et al., 2020 (link)) and measured by comparing binding to that of a tracer consisting of RvD1 linked to acetylcholinesterase (AChE) per the manufacturer’s protocol (Cayman Chemicals). The samples and the competition RvD1 tracer were incubated overnight at 4°C. The signal in ELISA plate was developed by Ellman’s Reagent, a substrate of AChE. The optical densities of samples were measured using an ELISA plate reader (Bio-Rad) at a wavelength of 420 nm and RvD1 levels were calculated using the standard curves.
3
Anti-inflammatory Effects of Platycodin D
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Platycodin D (purity >99%) was purchased from National Institutes for Food and Drug Control (Beijing, China). LPS (Escherichia coli O55:B5) and MTT were purchased from Sigma (St. Louis, MO, USA). TNF-α, IL-6, and IL-1β ELISA kits were purchased from Biolegend (CA, USA). Rabbit anti-human TLR4, NF-κB p65, IκBα, and β-actin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Rabbit anti-human LXRα and ABCA1 antibodies were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).
4
Cytokine Secretion in Podocytes
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Podocytes were grown on 35 mm collagen coated dishes as described above. Cells were exposed to culture media alone (control), 50 µM chloroquine alone in media, 5 mg/ml low endotoxin human albumin alone in media, or 50 µM chloroquine and 5 mg/ml low endotoxin albumin in media for 1, 3, or 18 hours. At the time of harvest, the cell supernatant was collected and the amount of cytokine present was analyzed using IL-6, TNF, or IL-1β ELISA kits (Biolegend, San Diego, CA). The podocytes in each dish were harvested in ice-cold (RIPA) buffer (50 mM Tris HCl, pH 8, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS) with Mini-Complete protease inhibitors added (Roche). Cytokine amount was normalized to total protein content as determined by the BCA assay.
5
Geraniin Inhibits Inflammatory Responses
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Geraniin (purity > 98%) was purchased from the Chinese Institute for Drug and Biological Product Control (Beijing, China). LPS was purchased from Sigma Chemical Co. (St. Louis, MO, USA). Mouse TNF-α,IL-6 and IL-1βELISA kits were obtained from BioLegend (CA, USA). Antibodies for Nrf2, HO-1, NF-κB p65, IκBα, and β-actin were purchased from CST (Danvers, MA, USA).
6
Quantifying Secreted IL-1β from Macrophages
7
Cytokine Quantification by ELISA
8
Quantification of Cytokines and Lipoprotein(a)
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The cultured supernatant from monocyte- and macrophage-treated H1/ H5 was harvested, and we performed ELISA. The cytokines level was determined by IL-1β ELISA Kit (BioLegend, San Diego, CA, USA) and IL-8 ELISA Kit (BD Biosciences, San Diego, CA, USA).
Serum levels of Lp(a) were determined using ELISA according to the manufacturer’s instruction (Mercodia, Uppsala, Sweden). The optical density at 450 nm was determined using a microplate absorbance reader Infinite M1000 Pro (TECAN, Männedorf, Switzerland). The data were analyzed with four parameters logistic curve on Myassays web site (https://www.myassays.com/ assessed on 8 March 2021). The unit of Lp(a) was presented as U/L; one U/L is equal to 1.254 mg/L.
Serum levels of Lp(a) were determined using ELISA according to the manufacturer’s instruction (Mercodia, Uppsala, Sweden). The optical density at 450 nm was determined using a microplate absorbance reader Infinite M1000 Pro (TECAN, Männedorf, Switzerland). The data were analyzed with four parameters logistic curve on Myassays web site (
9
Quantification of Secreted IL-1β
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Following specific treatments, the cell culture supernatants were collected, and the quantification of secreted IL-1β was performed using an IL-1β ELISA kit according to the manufacturer’s instructions (Biolegend, San Diego, CA, USA). Absorbance was measured using an ELISA reader at 450 nm.
10
Zoledronate and Bone Inflammation
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Zoledronate (Zol) was purchased from Sigma. An anti-OCN antibody was purchased from Abclonal. Anti-IL-1β, anti-TNF-α, and anti-IL-1RA antibodies were acquired from Abcam. Alexa Fluor 488 secondary antibody was purchased from Abcam. IHC-secondary antibodies were bought from ZSGB-BIO. IL-1 RA neutralizing and negative control antibodies were purchased from R&D Systems. IL-1β ELISA kit was purchased from Biolegend. Masson’s trichrome stain kit was obtained from Solarbio. Tartrate-resistant acid phosphatase (TRAP) stain kit was purchased from Sigma-Aldrich. Cell counting kit-8 was purchased from Dojindo.
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