Il 1β

Manufactured by Santa Cruz Biotechnology

Sourced in United States, United Kingdom, Canada

IL-1β is a recombinant human interleukin-1 beta protein. Interleukin-1 beta is a proinflammatory cytokine that plays a central role in the regulation of immune and inflammatory responses.

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Mouse anti-IL-1β (12 citations) IL-1β (sc-7884) (4 citations) Pro-IL-1β (3 citations) IL-1β (SC-12742) (3 citations) Anti–IL-1β (sc-52012, (3 citations) Anti-IL-1β (SC-12742), (3 citations) Anti-pro-IL-1β (3 citations) Interleukin (IL)-1β (2 citations) Rabbit anti-IL-1β (sc-7884) (2 citations) Mouse monoclonal anti-IL-1β (2 citations)

163 protocols using il 1β

Intestinal Inflammatory Protein Analysis

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Immunoblotting was performed as described previously [16 (link)]. Briefly, dissected intestinal were homogenized and lysed with lysis buffer. Protein extract was separated on SDS-PAGE and transferred to polyvinylidenedifluoride (PVDF) membrane and probed with the following antibodies: IL-6, IL-1β, TNF-α, PGE2, COX-2, and β-actin from Santa Cruz Biotechnology (Santa Cruz, CA). Band intensity was determined by densitometry analysis using the ImageJ analysis system.

Park S.A., Lee G.H., Hoang T.H., Lee H.Y., Kang I.Y., Chung M.J., Jin J.S, & Chae H.J. (2021). Heat-inactivated Lactobacillus plantarum nF1 promotes intestinal health in Loperamide-induced constipation rats. PLoS ONE, 16(4), e0250354.

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BDNF Signaling Protein Detection

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BDNF, leupeptin, aprotinin, phenyl-methylsulfonyl fluoride, pepstatin A, soybean trypsin inhibitor, NaF, sodium vanadate, glycerophosphate, 2-mercaptoethanol, NMDA, glycine, Tween 20, NP-40, and Histopaque-1077 were from Sigma. Anti-TrkB (SC-8316), -pY-Trk (SC-8058), -phospholipase C-γ1 (SC-7290), -N-Shc (SC-365598), -NR1 (SC-9058), -TNFα (SC-52746), -IL-6 (SC-57315), -IL-1β (SC-57315), -Arc (SC-365736), - BDNF/proBDNF (SC-2098), –NT3 (SC-547), -NT4 (SC-545), -β-actin (SC-47778) were from Santa Cruz Biotechnology. FNDC-5/irisin (36–335) was from ProSci (Poway, CA, USA). Seize-X immunoprecipitation kit, antigen elution buffer, Bind NeutrAvidin, high binding capacity coated 96-well plates, and West Pico chemiluminescent reagents were from Pierce-Endogen (Rockford, IL, USA). Bradford reagent, SDS-PAGE reagents, and pre-stained molecular weight markers were from Bio-Rad. Protease inhibitors (EDTA-free) and protein phosphatase inhibitor tablets were from Roche (Basel, Switzerland). BDNF was reconstituted according to the manufacturer’s instruction. To avoid freezing damage, 10% glycerol was added for a final concentration of 10 ng/μl BDNF and stored in 80°C until use. All other test agents were made fresh according to the manufacturer’s recommendation. The DMSO concentration in the incubation medium was 1%.

Meade G.M., Charron L.S., Kilburn L.W., Pei Z., Wang H.Y, & Robinson S. (2020). A Model of Negative Emotional Contagion Between Male-Female Rat Dyads: Effects of Voluntary Exercise on Stress-Induced Behavior and BDNF-TrkB Signaling. Physiology & behavior, 234, 113286.

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Western Blot Analysis of Inflammatory Proteins

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The collected cells were lysed in RIPA lysis solution containing 2 mM PMSF, and whole-cell proteins extracted and quantitated by the BCA assay (all protein extraction and quantification reagents were from Beyotime Biotechnology). Sixty μg of protein were then subjected to SDS-PAGE electrophoresis (Bio-rad), transferred to polyvinylidene fluoride (PVDF) membranes (Bio-Rad) and probed with primary antibodies followed by HRP-conjugated secondary antibodies. The immunoreactive bands were visualized with HRP substrate (Luminata, Millipore) using ChemiDocTM XRS₊ system (Bio-Rad). The following antibodies were used: ASC (sc-22514, Santa Cruz); Caspase-1 (sc-56036, Santa Cruz); IL-1β (sc-7884, Santa Cruz); NLRP3 (ab4207, Abcam); FXR (sc-1204, Santa Cruz); GAPDH (5174, CST); β-actin (8457L, CST); α/β-Tublin (2148, CST); anti-rabbit, anti-mouse HRP-conjugated second antibodies (7074S and 7076S, CST), and anti-goat HRP-conjugated second antibody (ab6885, Abcam).

Hao H., Cao L., Jiang C., Che Y., Zhang S., Takahashi S., Wang G, & Gonzalez F.J. (2017). Farnesoid X Receptor Regulation of the NLRP3 Inflammasome Underlies Cholestasis-Associated Sepsis. Cell metabolism, 25(4), 856-867.e5.

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Quantifying Inflammatory Markers in PVN

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The brain was sectioned serially in 300 μm increments from the bregma to lambda, both sides of the PVN tissues were isolated by the use of a punch-out technique with a cryostat [33 (link), 34 (link)], and the PVN tissue was stored at − 80 °C until use. Western blotting analysis was performed in the same manner as previously described [6 (link)]. The protein levels were determined from tissue homogenate obtained from the PVN for the following antibodies: NLRP3 (1:2000, Santa Cruz, CA, USA), ASC (1:500, Santa Cruz, CA, USA), pro-caspase-1 (1:2000, Abcam, MA, USA), IL-1β (1:500, Santa Cruz, CA, USA), CXCR3 (1:2000, Abcam, MA, USA), VCAM-1, ICAM-1 (1:2000, Abcam, MA, USA), and CCL2 (1:2000, Santa Cruz, CA, USA), Iba-1 (1:500, Santa Cruz, CA, USA). The β-actin antibody was used as an internal standard, and band densities were analyzed with NIH ImageJ software.

Wang M.L., Kang Y.M., Li X.G., Su Q., Li H.B., Liu K.L., Fu L.Y., Saahene R.O., Li Y., Tan H, & Yu X.J. (2018). Central blockade of NLRP3 reduces blood pressure via regulating inflammation microenvironment and neurohormonal excitation in salt-induced prehypertensive rats. Journal of Neuroinflammation, 15, 95.

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Immunohistochemical Analysis of Tmem43 Knockout Mice

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Hearts, kidneys and spleens from wild type and Tmem43 KI mice were collected and fixed with 4% paraformaldehyde overnight at 4 °C. Hearts and kidneys were embedded in paraffin, and cut into 4 μm sections. The sections were stained with Masson’s Trichrome staining (Polysciences Inc.) following the manufacturer’s protocol. The paraffin sections of mouse hearts or spleens were immunostained with specific antibodies: p65, p50, IL-1β, MAC2, TNFα, CD45 (Santa Cruz) and αSMA (Abcam, Cat# ab5694).

Zheng G., Jiang C., Li Y., Yang D., Ma Y., Zhang B., Li X., Zhang P., Hu X., Zhao X., Du J, & Lin X. (2018). TMEM43-S358L mutation enhances NF-κB-TGFβ signal cascade in arrhythmogenic right ventricular dysplasia/cardiomyopathy. Protein & Cell, 10(2), 104-119.

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Optimizing Celecoxib-Mangiferin Synergy Assessment

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MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), Tris, Ethylene diamine tetra acetic acid (EDTA), Trypsin-EDTA, Di-ethyl dithio-carbamate (DDC), Tween-20, Hematin, Glycerol, Phenol, Ammonium sulphate, Carrageenan, COX-1 and COX-2 enzymes were purchased from Sigma-Aldrich (St. Louis, MO, USA). Celecoxib was a generous gift from Unichem Laboratories (Mumbai, India). The stock solutions of PT-Mangiferin (100 mg/ml) was prepared in dimethyl sulphoxide and further dilutions were made in PBS for treatment in vitro and in vivo. Polyclonal antibodies to, COX-2, IL-1β and TNF-α were purchased from Santa Cruz Biotechnology (California, U.S.A.). iNOS antibody was from Thermo scientific. All other chemicals and solvents were of analytical grade and purchased from authorized standard companies.

Bulugonda R.K., kumar K.A., Gangappa D., Beeda H., Philip G.H., Muralidhara Rao D, & Faisal S.M. (2017). Mangiferin from Pueraria tuberosa reduces inflammation via inactivation of NLRP3 inflammasome. Scientific Reports, 7, 42683.

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NLRP3 Inflammasome Activation Assay

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Lipopolysaccharide (LPS; Escherichia coli 0111:B4), ATP, and oxidized ATP (oxATP) were obtained from Sigma (St. Louis, MO, USA), and nigericin was purchased from InvivoGen (San Diego, CA, USA). Ac-YVAD-cmk and MCC950(CP-456773 sodium salt) were purchased from Sigma (St. Louis, MO, USA). Cell Counting Kit-8 (CCK-8) was purchased from Dojindo (Japan). Calcein AM iwas purchased from Invitrogen (Carlsbad, CA, USA) and 1 mg/ml Propidium iodide solution was purchased from Sigma (St. Louis, MO, USA). 1% Crystal violet solution was purchased from Sigma (St. Louis, MO, USA). NLRP3 antibodies and secondary anti-sheep antibody conjugated to horseradish peroxidase (HRP) were purchased from R&D Systems (Minneapolis, MN, USA). Toll-like receptor 4 (TLR4), P2X7, ASC, IL-1β, and caspase-1 donkey anti-rabbit IgG-HRP were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Bae J.Y., Lee S.W., Shin Y.H., Lee J.H., Jahng J.W, & Park K. (2017). P2X7 receptor and NLRP3 inflammasome activation in head and neck cancer. Oncotarget, 8(30), 48972-48982.

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Gingival Expression of Inflammatory Markers

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Western blot analysis was utilized to evaluate the expression of IL-1β, IL-6 and ICAM-1 in mouse gingival tissues. Briefly, palatal gingival tissues were collected (2.0 mm × 2.5 mm) from mice in all the three groups 2 h after transfection. Total protein was then extracted from each tissue using RIPA buffer (Thermo Fisher Scientific, Waltham, MA, USA) and soluble proteins were resolved via 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride membrane (Immobilon-P, Millipore, Darmstadt, Germany). The blots were subsequently probed with a primary antibody for IL-6 (1:1 000; Novus Biologicals, Littleton, CO, USA), IL-1β (1:1 000; Santa Cruz Biotechnology, Dallas, TX, USA) or ICAM-1 (1:1 000; Proteintech, Chicago, IL, USA). Equal protein loading was confirmed by probing for β-actin(1:2 000; Santa Cruz Biotechnology). For chemiluminescence detection, we used a peroxidase-conjugated anti-rabbit IgG secondary antibody (1:10 000; Santa Cruz Biotechnology). The band intensities were measured using a ChemiDoc image analysis system (Fuji Film, Tokyo, Japan).

Yamaguchi H., Ishida Y., Hosomichi J., Suzuki J.I., Usumi-Fujita R., Shimizu Y., Kaneko S, & Ono T. (2017). A new approach to transfect NF-κB decoy oligodeoxynucleotides into the periodontal tissue using the ultrasound-microbubble method. International Journal of Oral Science, 9(2), 80-86.

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Western Blot Analysis of Signaling Proteins

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The proteins were extracted from the cell pellets. Proteins were boiled for 10 min and loaded onto 4–12% bis Tris gels (Invitrogen, Waltham, MA, USA) for separation. The separated proteins were then blotted onto polyvinylidene difluoride (PVDF) membranes (Invitrogen) with 20% (v/v) methanol in NuPage Transfer Buffer (Invitrogen) at 15 V for 4 h at 4°C. Tris-buffered saline containing 0.01% Tween 20 (TBST) in 5% skim milk (Difco, BD Biosciences, Oxford, UK) was used to block the membranes for 1 h. The blots were washed three times with TBST for 10 min and then incubated overnight at 4 °C with primary antibodies specific to the following target proteins: phosphorylated JAK1, phosphorylated JAK2, phosphorylated STAT1 (1:1000; Cell Signaling Technology, Cambridge, UK), CSF1, IL-6, PTPN6, RAC2, TNFα, IL-1α, IL-1β, phosphorylated STAT3, STAT3 ADORA2A, JAK1, JAK2, STAT1, SOCS3, Bax, Bcl-2, and β-actin (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA). After incubation, the blots were washed thrice with TBST and incubated for 1 h with a horse-radish peroxidase–conjugated secondary antibody (1:3000; Santa Cruz) at 25°C. Finally, the blots were visualized using an enhanced chemiluminescence detection system (Amersham Pharmacia Biotech, Little Chalfont, UK).

Jo S., Baek A., Cho Y., Kim S.H., Baek D., Hwang J., Cho S.R, & Kim H.J. (2023). Therapeutic effects of polydeoxyribonucleotide in an in vitro neuronal model of ischemia/reperfusion injury. Scientific Reports, 13, 6004.

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Oesophageal Tissue Protein Extraction and Western Blotting

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Oesophageal strips or cultured cells were collected to extract the protein with RIPA lysis buffer (Beyotime Biotech), which contained 1 mmol/L final concentration of phenylmethanesulfonyl fluoride (PMSF). After complete lysis, the samples were centrifuged at 10 000 g for 5 minutes at 4°C to precipitate the tissue debris. The supernatants were used to measure the protein concentration by a BCA Protein Assay Kit (Beyotime Biotech). The proteins were electrophoresed in 10% SDS‐PAGE gels and then transferred to PVDF membranes. After blocking with 5% skim milk for 1 hour at room temperature, the membranes were incubated with the following primary antibodies: CaSR (1:200, ab19347, Abcam), NLRP3 (1:100, ab214185, Abcam), ASC (1:100, sc‐22514‐R, Santa Cruz), Caspase‐1 p20 (1:100, sc‐398715, Santa Cruz) and IL‐1β (1:100, sc‐32294, Santa Cruz) at 4°C overnight. The membrane was washed with TBST and incubated with secondary antibodies for 2 hours at room temperature. Protein bands were visualized on the membrane with a Gel Imaging System, and the protein bands were quantified with ImageJ software.

Yin X., Wu H., Zhang B., Zhu N., Chen T., Ma X., Zhang L., Lv L., Zhang M., Wang F, & Tang X. (2019). Tojapride prevents CaSR‐mediated NLRP3 inflammasome activation in oesophageal epithelium irritated by acidic bile salts. Journal of Cellular and Molecular Medicine, 24(2), 1208-1219.

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