Ab232898

Manufactured by Abcam

Sourced in United Kingdom, United States

Ab232898 is a lab equipment product manufactured by Abcam. It is designed for specific research applications, but a detailed description cannot be provided while maintaining an unbiased and factual approach. Further information about the core function of this product is not available.

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Lab products found in correlation

Ab126706, by Abcam (3 mentions) Leupeptin, by Selleck Chemicals (1 mentions) Hsp90, by Santa Cruz Biotechnology (1 mentions) Protease and phosphatase inhibitor, by Roche (1 mentions) Protease inhibitor phenylmethanesulfonyl fluoride, by Beyotime (1 mentions) Ecl system, by Beyotime (1 mentions) 0.2 μm pvdf membrane, by Merck Group (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Ab8227, by Abcam (1 mentions) Bradford protein assay kit, by Beyotime (1 mentions) Ab32053, by Abcam (1 mentions) Ab32503, by Abcam (1 mentions) Ab32124, by Abcam (1 mentions) Radioimmunoprecipitation assay buffer, by Applygen (1 mentions) Ab137550, by Abcam (1 mentions) Ab39969, by Abcam (1 mentions)

5 protocols using ab232898

Analyzing Autophagy and Lipid Metabolism

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We lysed mouse tissues or cells in 2% SDS)buffer supplemented with phosphatase and protease inhibitors (Roche). We used a BCA assay (Pierce, Thermo Fisher) to test total protein concentration. As a measure of autophagic flow, Western blots for LC3 and p62 were performed in cells treated with the lysosomal inhibitor, leupeptin (Selleck;10 μM) for 12 h. We loaded 30 μg of protein to SDS-PAGE, and subsequently transferred the protein onto 0.22 μm PVDF membranes. We used 5% BSA to block membranes and then incubated the membranes with primary antibodies and respective secondary antibodies and subjected them to electrochemiluminescence.
The primary antibodies used in this experiment were: p-HSL^ser660 (1:1,000; CST, CST4139), total-HSL (1:1,000; CST, CST4107), HSP90 (1:1,000; Santa Cruz, sc7947), ATGL (1:1,000; CST, CST2138), SCD1 (1:1,000; CST, CST2794), p62 (1:1,000; MBL, PM045), LC3 (1:1,000; MBL, M186-3), FA desaturase-1 [FADS1 (delta-5-desaturase)] (1:1,000; Abcam, ab126706), FA desaturase-2 [FADS2 (delta-6-desaturase)] (1:1,000; Abcam, ab232898).

Zou Y., Wang Y.N., Ma H., He Z.H., Tang Y., Guo L., Liu Y., Ding M., Qian S.W, & Tang Q.Q. (2020). SCD1 promotes lipid mobilization in subcutaneous white adipose tissue. Journal of Lipid Research, 61(12), 1589-1604.

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Western Blotting for Protein Analysis

Cited 1 time

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Western blotting was performed as described previously (10 (link)). Briefly, to obtain total protein, cells or tissues were lysed with RIPA lysis buffer (Beyotime, Shanghai, China) containing phenylmethanesulfonyl fluoride protease inhibitor (Beyotime). Protein concentration was measured using a detergent compatible Bradford protein assay kit (Beyotime). In total, 20 μg protein was loaded onto a 10% denaturing SDS-PAGE gel, and protein bands were then transferred to a 0.2-μm PVDF membrane (Merck Millipore, Germany). The membrane was blocked in 3% bovine serum albumin for 1 h, and FADS2 rabbit polyclonal antibody (Abcam, ab232898, 1:1000 dilution), E-cadherin rabbit polyclonal antibody (Proteintech, 20874-1-AP, 1:5000 dilution), N-cadherin rabbit polyclonal antibody (Proteintech, 66219-1-Ig, 1:5000 dilution), Snail rabbit polyclonal antibody (Proteintech, 13099-1-AP, 1:500 dilution), GAPDH rabbit polyclonal antibody (Proteintech, 10494-1-AP, 1:20000 dilution), or β-Actin rabbit polyclonal antibody (Abcam, ab8227, 1:5000 dilution) was added, followed by incubation at 4°C. After overnight incubation, the membrane was incubated with rabbit (Epizyme, LF102, 1:5,000 dilution) secondary antibody. Finally, the proteins bands were detected using the ECL system (Beyotime Biotechnology, China). This experiment was performed at least three times.

Zhao T., Gao P., Li Y., Tian H., Ma D., Sun N., Chen C., Zhang Y, & Qi X. (2023). Investigating the role of FADS family members in breast cancer based on bioinformatic analysis and experimental validation. Frontiers in Immunology, 14, 1074242.

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Quantification of Protein Expression in Macrophages and Tumor Cells

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Protein from macrophages, THP-1 cells, FaDu cells, and tumor tissues were dissolved in radioimmunoprecipitation assay buffer (Applygen, Beijing, China). Equal amounts of protein lysates were loaded onto a 10% SDS-PAGE gel for electrophoresis and transferred to a PVDF membrane. After blocking with 5% nonfat milk, the membranes were incubated overnight at 4 °C with primary antibodies against FADS1 (ab126706, Abcam, UK), FADS2 (ab232898, Abcam, UK), cyclin B1 (ab32053, Abcam, UK), CDK1 (ab210008, Abcam, UK), Bcl-2 (ab32124, Abcam, UK), and BAX (ab32503, Abcam, UK). The next day, the membranes were then incubated with secondary antibodies (dilution 1:5000) for 1 h at 25 °C. After washing three times with TBST for 10 min, the blots were visualized using a chemiluminescence western blotting system and quantified using ImageJ software (version 1.8.0, National Institutes of Health, USA).

Gong Q., Li H., Song J, & Lin C. (2023). LncRNA LINC01569 promotes M2 macrophage polarization to accelerate hypopharyngeal carcinoma progression through the miR-193a-5p/FADS1 signaling axis. Journal of Cancer, 14(9), 1673-1688.

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Immunohistochemical Analysis of Esophageal Lesions

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Immunohistochemical staining was performed on formalin‐fixed paraffin‐embedded tissues in a tissue microarray format, using antibodies against DEGS1 (Abcam#ab167169), FADS1 (Abcam#ab236672), FADS2 (Abcam#ab232898) and Nrf2 (Abcam#ab137550). Staining details are provided in Supporting Information. Staining intensities in GC, NSE, BE, LGD, HGD, and intraepithelial carcinoma (IEC) were scored by a specialist gastrointestinal pathologist. The phenotype was determined on the basis of H&E‐stained sections, according to the standard diagnostic procedure. Phenotypes occasionally changed between sequential sections, such that slightly different numbers exist for each analysis. Scoring was based on the phenotype in the region of interest: for example, NSE refers to only the normal squamous epithelium in the section: this is in contrast to the omics data which use the whole biopsy, where normal esophageal tissue may include stroma, submucosal glands, as well as epithelium. Each component, if present in the tissue, was scored separately using a 4‐grade assessment of intensity (0 no staining, 1+ weak staining, 2+ moderate staining, 3+ strong staining). Where the staining was non‐uniform in a component, the maximum intensity of staining was used for the score, providing at least 10% of the cells of that component stained to this intensity.

Molendijk J., Kolka C.M., Cairns H., Brosda S., Mohamed A., Shah A.K., Brown I., Hodson M.P., Hennessy T., Liu G., Stoll T., Richards R.S., Gartside M., Patel K., Clemons N.J., Phillips W.A., Barbour A., Westerhuis J.A, & Hill M.M. (2022). Elevation of fatty acid desaturase 2 in esophageal adenocarcinoma increases polyunsaturated lipids and may exacerbate bile acid‐induced DNA damage. Clinical and Translational Medicine, 12(5), e810.

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Immunofluorescence Analysis of FADS Enzymes

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Media were aspirated and cells rinsed twice in 100 μl PBS, fixed and permeabilized in 70% acetone and 30%methanol at −20 °C for 10 min, air-dried, and incubated in blocking solution (10% Normal Donkey Serum in PBS) for 1 h at room temperature. Primary antibodies against FADS1 (ab126706), FADS2 (ab232898)) and FADS5 (ab39969) were purchased from Abcam (Cambridge, MA, USA). Antibodies against the specific proteins, and respective isotype controls, were applied at concentrations of 4 μg/mL overnight at 4 °C. Antispecies anti-rabbit secondary antibodies conjugated to Alexa Fluor 555 were purchased from Molecular Probes and Thermo Fisher Scientific. Slides were washed three times for 10 min each in PBS to eliminate excess antibody, and secondary antibodies were applied at a 1:100 dilution, for 1 h at room temperature. Confocal images were collected using a Zeiss Observer.Z1 inverted microscope equipped with Semrock wheel filters with ec/em spectra for the Alexa Fluor probes used. Images were acquired and analyzed using Slidebook 6.0 (Intelligent Imaging Innovations, Denver, CO).

Thomas T., Cendali F., Fu X., Gamboni F., Morrison E.J., Beirne J., Nemkov T., Antonelou M.H., Kriebardis A., Welsby I., Hay A., Dziewulska K.H., Busch M.P., Kleinman S., Buehler P.W., Spitalnik S.L., Zimring J.C, & D’Alessandro A. (2021). Fatty acid desaturase activity in mature red blood cells and implications for blood storage quality. Transfusion, 61(6), 1867-1883.

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