Akr1c3

Manufactured by Abcam

Sourced in United States, Canada

AKR1C3 is an enzyme that is involved in the metabolism of various steroid hormones and xenobiotic compounds. It catalyzes the reduction of a wide range of carbonyl-containing compounds, including steroids, prostaglandins, and other xenobiotics. AKR1C3 plays a role in the regulation of androgen and estrogen levels in the body.

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Lab products found in correlation

Ripa buffer, by Thermo Fisher Scientific (2 mentions) Anti goat horseradish peroxidase hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Akr1c1, by Abcam (1 mentions) Txnrd1, by Abcam (1 mentions) Anti rabbit and anti mouse hrp conjugated secondary antibodies, by Cell Signaling Technology (1 mentions) L glutamine, by Nacalai Tesque (1 mentions) Mem nonessential amino acid solution, by Nacalai Tesque (1 mentions) Rabbit igg, by Santa Cruz Biotechnology (1 mentions) Fetal bovine serum (fbs), by Equitech-Bio (1 mentions) Bca protein assay kit, by Abcam (1 mentions) β actin, by Abcam (1 mentions) Srebp1, by Santa Cruz Biotechnology (1 mentions) Sema3c, by Santa Cruz Biotechnology (1 mentions) Anti mouse alexa fluor 680, by Thermo Fisher Scientific (1 mentions) Alexa fluor anti rabbit 680, by Thermo Fisher Scientific (1 mentions) Anti goat alexa fluor 680, by Thermo Fisher Scientific (1 mentions) Hmgcs1, by Cell Signaling Technology (1 mentions) Odyssey system, by LI COR (1 mentions) Actin, by Merck Group (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions)

3 protocols using akr1c3

Purification and Evaluation of Isothiocyanates

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ITCs (SFN, 6-MSITC, and 6-MTITC) were purified from Wasabi by reversed-phase high performance liquid chromatography (HPLC) to >99.3% purity26 (link) and dissolved in dimethyl sulfoxide for cell culture experiments. The antibodies against Nrf2 (C-20), Keap1 (E-20), NQO1 (C-19), HSP70 (D69), GAPDH, rabbit IgG, and horseradish peroxidase (HRP)-conjugated anti-goat secondary antibody were purchased from Santa Cruz Biotechnology. AKR1C1, AKR1C3, and TXNRD1 antibodies were obtained from Abcam. HRP-conjugated anti-rabbit and anti-mouse secondary antibodies were from Cell Signaling Technology.
IMR-32 cell culture. Human neuroblastoma IMR-32 cells (cell no. TKG0207) were obtained from Riken Bioresource Center Cell Bank. IMR-32 cells were grown in Eagle’s Minimum Essential Medium (Nissui Seiyaku) supplemented with 2 mM l-glutamine (Nacalai Tesque), 1% v/v MEM nonessential amino acid solution (Nacalai Tesque), and 10% v/v fetal bovine serum (Equitech-Bio) under a humidified 5% CO₂ atmosphere at 37 °C.

Trio P.Z., Fujisaki S., Tanigawa S., Hisanaga A., Sakao K, & Hou D.X. (2016). DNA Microarray Highlights Nrf2-Mediated Neuron Protection Targeted by Wasabi-Derived Isothiocyanates in IMR-32 Cells. Gene Regulation and Systems Biology, 10, 73-83.

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Quantitative Western Blot Analysis

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Total protein was obtained from the lysed cells with RIPA buffer (Thermo Scientific, USA), and the protein concentration was quantified with BCA Protein Assay Kit (Abcam, USA). Protein electrophoresis and transferring to nitrocellulose membrane followed the routine protocols described in our previous research. The membrane was blocked with bovine serum albumin (5%) for 1 h, and the membrane was hatched overnight with the primary antibodies against HOXB4, AKR1C3, GPX4, and β-actin (Abcam, USA). The membrane was washed with TBST for five times and incubated with the second antibodies at 37°C for 1 h. Enhanced Chemiluminescence Reaction Solution (Pierce, Rockford, IL, USA) was added to the membrane for 1 min. We chose β-actin as the internal reference. The protein images were explicated with ImageJ2x, and Gelpro 32 software was used to analyze the gray values.

Liang J., Cao Y., He M., Li W., Huang G., Ma T., Li M., Huang Y., Huang X, & Hu Y. (2021). AKR1C3 and Its Transcription Factor HOXB4 Are Promising Diagnostic Biomarkers for Acute Myocardial Infarction. Frontiers in Cardiovascular Medicine, 8, 694238.

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Western Blot Analysis of Prostate Cancer Markers

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Lysis buffer contained 50 mM Tris-HCl, 150 mM NaCl, 1% NP40, 10 mM NaF, 10% Glycerol, and protease inhibitor cocktail (Roche, ON, Canada. Cat. No. 04693116001). Western blots were imaged by a LI-COR Odyssey system. Loading controls were run on the same blots. Primary antibodies include: AR (Santa Cruz Biotechnology, TX, USA. Cat. No. sc-816), PSA (Cell Signaling Technology, MA, USA. Cat. No. 5877), SEMA3C (Santa Cruz Biotechnology Cat. No. sc-27796), AKR1C3 (Abcam, ON, Canada. Cat. No. ab49680), HSD3B2 (Abnova, ON, Canada. Cat. No. 3284-mo2), HSD17B3 (Abnova Cat. No. 29–119), FASN (Cell Signaling Technology. Cat. No. 3180), HMGCS1 (Cell Signaling Technology. Cat. No. 36877), SREBP-1 (Santa Cruz Biotechnology. Cat. No. sc-13551), UGT2B17 (Abcam. Cat. No. ab126269) and actin (Sigma-Aldrich, ON, Canada. Cat. No. A2066). Secondary antibodies were anti-rabbit alexa fluor 680 (Invitrogen, Cat. No. A21109), anti-mouse alexa fluor 680 (Invitrogen, Cat. No. A21058), anti-goat alexa fluor 680 (Invitrogen, Cat. No. A21084).

Battin T.J., Wille A., Sattler B, & Psenner R. (2001). Phylogenetic and Functional Heterogeneity of Sediment Biofilms along Environmental Gradients in a Glacial Stream. Applied and Environmental Microbiology, 67(2), 799-807.

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