Ab207802

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab207802 is a primary antibody developed for use in immunohistochemistry (IHC) applications. It targets an unspecified antigen. The antibody is produced in rabbit and is available in liquid format.

Automatically generated - may contain errors

Lab products found in correlation

Ab263899, by Abcam (16 mentions) Ab215203, by Abcam (10 mentions) Ab214185, by Abcam (8 mentions) Pvdf membrane, by Merck Group (7 mentions) Ab181602, by Abcam (6 mentions) Ab155970, by Abcam (5 mentions) Ab254360, by Abcam (5 mentions) Ab16502, by Abcam (4 mentions) Ab9485, by Abcam (4 mentions) Ab180673, by Abcam (3 mentions) Ripa lysis buffer, by Solarbio (3 mentions) Ab8245, by Abcam (3 mentions) Ab8227, by Abcam (3 mentions) Ab219800, by Abcam (3 mentions) Bca protein assay kit, by Beyotime (3 mentions) Polyvinylidene fluoride membrane, by Merck Group (3 mentions) Ab138483, by Abcam (2 mentions) Ab22684, by Abcam (2 mentions) Ab238124, by Abcam (2 mentions) Ab76302, by Abcam (2 mentions)

Close spelling variants of this product

Caspase-1 (ab207802), (2 citations)

42 protocols using ab207802

NLRP1 Activation and Autophagy Signaling

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The NLRP1 activator muramyl dipeptide (MDP) (Selleck, USA) was then added at 100 μmol for 24 hours to the oxidative stress model according to the preliminary experiment and the published studies [21 (link)–24 (link)]. The cell viability determination was the same as that in Section 2.1.
Total protein from HTR-8/SVneo cells was prepared with RIPA lysing buffer. The sample proteins (20 μg) of the different groups were separated using 10% SDS-PAGE and transferred onto PVDF membranes. These membranes were incubated with a primary antibody, followed by incubation of the anti-rabbit IgG secondary antibody. Protein expression was detected with an enhanced chemiluminescence detection kit. GAPDH served as an internal control. The antibodies included those for IL-1β (Abcam, ab216995), pro-IL-1β (Abcam, ab216995), pro-CASP1 (Abcam, ab207802), CASP1 (Abcam, ab207802), Beclin-1 (Abcam, ab207612), LC3 (Abcam, ab51520), p62 (Abcam, ab211324), ATG5 (Abcam, ab108327), ATG7 (Abcam, ab52472), NLRP3 (Abcam, ab263899), NLRP1 (BioLegend, 679802), and GAPDH (Abcam, ab128915). An ECL chemiluminescent reagent and imaging system (Bio-Rad, USA) were used to display protein bands, with the collected images analyzed using Bio-Rad software.

Li M., Sun T., Wu X., An P., Wu X, & Dang H. (2021). Autophagy in the HTR-8/SVneo Cell Oxidative Stress Model Is Associated with the NLRP1 Inflammasome. Oxidative Medicine and Cellular Longevity, 2021, 2353504.

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Western Blot Analysis of Apoptosis Markers

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RIPA (GBCBIO, G3424) was used to lyse cells and extract total protein to prepare protein samples. The protein lysates were separated by conventional electrophoresis, transferred onto the membrane, and washed with PBS, followed by blocking with 1% BSA for 2 h. Then, primary antibodies Box (ab32503, Abcam, Cambridge, UK), Bcl-2 (ab32124, Abcam, Cambridge, UK), Caspase-1 (ab32503, Abcam, Cambridge, UK), Cleaved Caspase-3 (ab32503, Abcam, Cambridge, UK), Caspase-8 (ab207802, Abcam, Cambridge, UK), Caspase-9 (ab2302, Abcam, Cambridge, UK), Caspase-11 (ab25901, Abcam, Cambridge, UK), Cyclin D1 (ab202068, Abcam, Cambridge, UK), ERK 1/2 (ab180673, Abcam, Cambridge, UK), p-ERK 1/2 (ab16663, Abcam, Cambridge, UK), WNT (ab17942, Abcam, Cambridge, UK), β-catenin (ab223500, Abcam, Cambridge, UK), GSDMD (ab15251, Abcam, Cambridge, UK) and β-actin (ab32572, Abcam, Cambridge, UK) (abcam, ab32503, ab32124, ab207802, ab2302, ab25901, ab202068, ab180673, ab16663, ab17942, ab223500, ab15251, ab32572, ab219800, ab8227) each at 1:1000 dilution were incubated with the membrane overnight at 4°C. The next day, HRP (ab32572, Abcam, Cambridge, UK) and labeled IgG secondary antibody (ab150077, Abcam, Cambridge, UK) each at 1:5,000 were further incubated with the membrane at room temperature for 1 h. ECL Luminescence Kit (Thermo, 32209) was used to develop the signal in a dark room.

He X., Li C., Yin H., Tan X., Yi J., Tian S., Wang Y, & Liu J. (2022). Mesenchymal stem cells inhibited the apoptosis of alveolar epithelial cells caused by ARDS through CXCL12/CXCR4 axis. Bioengineered, 13(4), 9060-9070.

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Western blotting of NLRP3 inflammasome

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The western blotting was performed as described previously [20 (link)]. The primary antibodies used in the present study were obtained from Abcam. Lysis buffer was used to prepare protein of cells. Same amounts of protein were loaded on an SDS-PAGE and then transferred electrophoretically to PVDF membrane (Millipore, USA). After blocking with TBST containing 5% milk, the membranes were incubated with primary antibody (1:1000) at 4 °C overnight. After washing and incubation, the membranes were incubated with secondary antibody (1:2000) in TBST. ECL Plus detection system (Millipore, USA) was applied for immuno-detection, e-Blot software was used for acquisition, and Image J software was used for analysis. The antibodies used in this study are listed as follows: rabbit monoclonal to NLRP3 (#ab263899, abcam, UK), rabbit polyclonal to ASC1 (#ab70627, abcam, UK), Rabbit monoclonal to caspase-1 (#ab207802, abcam, UK), Rabbit monoclonal to SIRP alpha (#ab302974, abcam, UK), Rabbit monoclonal to Connexin 43 (#ab214274, abcam, UK). The experiments were repeated at least 3 independent times. The original blots/gels are presented in Additional files 2, 3, 4: Figs. S2–S4, and the gels were cut prior to hybridisation with antibodies to save antibody, which is common in most labs. Therefore, we are unable to provided full-length blots/gels.

Wu H., Dai R., Wang M, & Chen C. (2023). Uric acid promotes myocardial infarction injury via activating pyrin domain-containing 3 inflammasome and reactive oxygen species/transient receptor potential melastatin 2/Ca2+pathway. BMC Cardiovascular Disorders, 23, 10.

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Western Blot Analysis of Caspase Activation

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The treated cells were lysed in RIPA buffer containing 1 mM DTT, 11 μg/ml DNase I, and protease inhibitor cocktail (Roche) and incubated on ice for 30 min. Sixty micrograms of protein sample were separated by electrophoresis on a denatured 10% SDS-PAGE gel and blotted onto a PVDF membrane (Millipore). The primary antibodies (Anti-Caspase-1, 1:1,000, ab138483, Abcam; Anti-cleavage Caspase-1, 1:1,000, ab207802, Abcam; Anti-Caspase-11, 1:1,000, ab22684, Abcam Anti-cleavage Caspase-11, 1:1,000, ab180673, Abcam) were incubated overnight at 4°C. Rabbit monoclonal to GAPDH (EPR16891, Abcam) was used as a loading control. Afterward, the membranes were rinsed and subsequently incubated with appropriate secondary antibodies. An ECL kit (Beyotime Biotechnology, Shanghai, China) was used to detect the bands of Western blots.

Wang J., Li X., Liu Y., Peng C., Zhu H., Tu G., Yu X, & Li Z. (2020). CircHIPK3 Promotes Pyroptosis in Acinar Cells Through Regulation of the miR-193a-5p/GSDMD Axis. Frontiers in Medicine, 7, 88.

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Western Blotting Standardized Protocols

Cited 1 time

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Western blotting was performed using the standard procedures as we previously reported^{58 (link),59 (link)}. Proteins were extracted from cells or tissues using RIPA lysis buffer (Solarbio). Equal volumes of lysates were loaded and separated on 10% SDS-PAGE gels and blotted on polyvinylidene difluoride membrane. After being blocked with 5% skim milk for 2 h at room temperature, the membranes were incubated with primary antibodies overnight at 4 ˚C. The primary antibody used in western blot analysis included Anti-ASC (1:1000; ab155970; Abcam); anti-caspase-1 (1:1000; ab207802; Abcam); anti-caspase-4 (1:1000; ab238124; Abcam); anti-GSDMD (1:1000; 66387-1-Ig; Proteintech); anti-PD1 (1:1000; ab52587; Abcam) and anti-GAPDH (1:500; ab8245; Abcam;). Anti-rabbit-HRP (1:5000; #7074, Cell Signaling Technology) and anti-mouse-HRP (1:5000; #7076, Cell Signaling Technology) were used as secondary antibodies and incubated at room temperature for 1 h. The blots were detected using a chemiluminescence kit (cat. no. 34577; Thermo Fisher Scientific, Inc.) and imaged using MiniChemi 610 system (Sage Creation Science, Co., Ltd.).

Lou X., Li K., Qian B., Li Y., Zhang D, & Cui W. (2022). Pyroptosis correlates with tumor immunity and prognosis. Communications Biology, 5, 917.

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Protein Extraction and Western Blot Analysis

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RIPA lysis buffer (Sigma, Shanghai, China) was applied to extract total proteins. A BCA kit (Sigma) was employed to measure the concentration of protein in the supernatant. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis was used to separate proteins, followed by their electrical transfer to a polyvinylidene fluoride membrane. Post blocking with 5% BSA, membranes were kept overnight at 4 °C alongside diluted primary antibodies: anti-USP11 (1:5000, ab109232, Abcam), anti-NLRP3 (1:1000, ab263899, Abcam, Cambridge, UK), anti-GSDMD-N (1:1000, ab215203, Abcam), anti-caspase-1 (1:1000, ab207802, Abcam), anti-IL-1β (1:1000, ab300501, Abcam), anti-IL-18 (1:1000, ab207323, Abcam), anti-p-IKKβ (1:1000, ab194528, Abcam), anti-IKKβ (1:500, ab32135, Abcam), anti-p-NF-kB (1:1000, ab76302, Abcam), anti-NF-kB (1:1000, ab16502, Abcam), anti-TRAF3 (1:1000, ab155298, Abcam), and anti-β-actin (1:1000, ab8227, Abcam) and then incubation for 1 h by secondary antibody. An ECL chromogenic substrate was used to visualize the bands.

Zhang Y., Hailati J., Ma X., Midilibieke H, & Liu Z. (2023). Ubiquitin-specific protease 11 Aggravates Ischemia-reperfusion-induced Cardiomyocyte Pyroptosis and Injury by Promoting TRAF3 Deubiquitination. Balkan Medical Journal, 40(3), 205-214.

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Western Blot Analysis of Inflammatory Signaling

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Protein samples (20 µg) were separated by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the proteins were then transferred into polyvinylidene fluoride blots. After being blocked by 5% milk, the blots were incubated with an antibody against target proteins at 4 °C overnight. Antibodies against p-NF-κB (CST, #3033) was obtained from Cell Signaling Technology (Danvers, MA). Antibodies against NF-κB antibody (ab16502), NLRP3 (ab214185), Caspase-1 (ab207802), IL-1β (ab229696), TRAF6 (ab40675), IRAK1 (ab238) were purchased from Abcam (Cambridge, UK). The α-Tubulin antibody (Sigma, T5168) was incubated as a loading control. These blots were further incubated with horseradish peroxidase (HRP)-conjugated secondary antibody (Abcam, ab6789 and ab6721), and exposed on a ChemiDoc MP imaging system (BIO-RAD) with an enhanced chemiluminescence (ECL) solution.

Hou J., Deng Q., Deng X., Zhong W., Liu S, & Zhong Z. (2021). MicroRNA-146a-5p alleviates lipopolysaccharide-induced NLRP3 inflammasome injury and pro-inflammatory cytokine production via the regulation of TRAF6 and IRAK1 in human umbilical vein endothelial cells (HUVECs). Annals of Translational Medicine, 9(18), 1433.

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NLRP3 and Caspase-1 Protein Expression

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BEAS-2B cells were treated under different conditions. The total protein was extracted from the cells using RIPA buffer (Abcam, Cambridge, UK) containing protease and phosphatase inhibitors (1 µg/ml Aprotinin, 1 µg/ml Leupeptin, 200 µM phenylmethylsulfonyl fluoride (PMSF), 1 mM Na₃VO₄). Then, the total protein was separated and transferred to membranes. The membranes were incubated with 5% non-fat milk and specific antibodies against NLRP3 (1:1000; ab263899, Abcam), caspase-1 (1:1000; ab207802, Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:1000; 2118, Cell Signaling Technology, Beverly, MA, USA). Finally, the proteins were visualized using ImageQuant LAS 4000 (GE Healthcare, Uppsala, Sweden) or X-ray film in a darkroom.

Chung C., Park S.Y., Huh J.Y., Kim N.H., Shon C., Oh E.Y., Park Y.J., Lee S.J., Kim H.C, & Lee S.W. (2024). Fine particulate matter aggravates smoking induced lung injury via NLRP3/caspase-1 pathway in COPD. Journal of Inflammation (London, England), 21, 13.

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Protein Expression Analysis in HK-2 Cells

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Total protein was extracted from HK-2 cells using RIPA lysis buffer containing protease inhibitor cocktail (Thermo Fisher Scientific, MA, USA). Cells suspended in RIPA buffer were lysed on ice for 10 min and the cell lysate was centrifuged at 12,000 × g for 10 min. Protein concentration was quantified by a BCA Protein Assay Kit (Beyotime Biotechnology, Shanghai, China). Fifteen micrograms of protein was used for SDS-PAGE electrophoresis, and separated proteins were transferred onto the PVDF membrane. After blocking with 5% skimmed milk for 1 h, the membrane was then incubated with primary antibodies (1:1000 dilution, all from Abcam), including NLRP3 (ab263899), Caspase-1 (ab207802), IL-1β (ab254360), GSDMD-N (ab215203), SGK1 (ab32374), and GAPDH (ab8245). After washing with TBS Tween 20, membranes were subject to further probing using HRP-linked secondary antibody (1:3000; Cell signaling technologies, MA, USA). An ECL chemiluminescence kit (Thermo Fisher Scientific, MA, USA) was employed to detect protein bands, and GAPDH was used for internal reference for protein quantification using ImageJ software (Bethesda, MD, USA).

Zhuang L., Jin G., qiong W., Ge X, & Pei X. (2023). Circular RNA COL1A2 Mediates High Glucose-Induced Oxidative Stress and Pyroptosis by Regulating MiR-424-5p/SGK1 in Diabetic Nephropathy. Applied Biochemistry and Biotechnology, 195(12), 7652-7667.

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Western Blot Analysis of Apoptosis Markers

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Protein expressions of caspase-1, Capase-4, and cleaved N-terminal gasdermin D (GSDMD-N) were measured by Western blotting [26 (link)]. Proteins were extracted from cells and tissues by RIPA (Sigma-Aldrich, Missouri, USA) reagents and were detected using the BCA method (P0012; Beyotime, Shanghai, China). Next, 30 μg total protein from each well was separated by 10% SDS-PAGE electrophoresis (P0690; Beyotime, Shanghai, China) and transferred to a PVDF membrane (FFP32; Beyotime, Shanghai, China). The membranes were blocked with 5% defatted milk for 1 h. Then, primary antibodies, including anti-caspase-1 (ab207802, 1/1000), anti-Capase-4 (ab238124, 1/1000), and anti-GSDMD-N (ab215203, 1/1000), were added (Abcam, California, USA). The next day, HRP labeled secondary antibody (ab7090, Abcam, California, USA) was added after the membrane was washed with PBS, and the membrane was incubated at room temperature for 1 h. The membranes were developed using ECL reagents with GAPDH as control.

Pan J., Zhao L., Liu J, & Wang G. (2022). Inhibition of circular RNA circ_0138959 alleviates pyroptosis of human gingival fibroblasts via the microRNA-527/caspase-5 axis. Bioengineered, 13(1), 1908-1920.

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