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Ncoa4

Manufactured by Cell Signaling Technology
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NCOA4 is a protein involved in cellular processes. It is a product used in laboratory research settings for the study of its biological functions.

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Lab products found in correlation

Slc7a11, by Cell Signaling Technology (2 mentions) Phenylmethanesulfonyl fluoride, by Solarbio (1 mentions) Phosphatase inhibitor cocktail, by Selleck Chemicals (1 mentions) Mmp13, by Abcam (1 mentions) Collagen 2, by Boster Bio (1 mentions) Mmp 3, by Cell Signaling Technology (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Ripa lysis buffer, by Solarbio (1 mentions) Cox 2, by Proteintech (1 mentions) Epcam, by Cell Signaling Technology (1 mentions) Phospho p38 mapk p p38 mapk, by Cell Signaling Technology (1 mentions) Caspase 3, by Cell Signaling Technology (1 mentions) α smooth muscle actin α sma, by Cell Signaling Technology (1 mentions) Histone h3, by Cell Signaling Technology (1 mentions) P38 mapk, by Cell Signaling Technology (1 mentions) Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions) Bcl 2, by Cell Signaling Technology (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions)

4 protocols using ncoa4

1

Protein Extraction and Western Blot Analysis of Human Cartilage

Cited 1 time
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The total protein from human cartilages and chondrocytes with various treatment were extracted using RIPA lysis buffer (#R0010, Solarbio) containing 1 mM phosphatase inhibitor cocktail (#B15002, Bimake, USA) and 1 mM phenylmethanesulfonyl fluoride (#329-98-6, Solarbio). Western blot was performed essentially as previously described.18 (link) The primary antibodies used were: NCOA4 (#66849, Cell Signaling Technology), Tf (#17435-1-AP, Proteintech, RRID:AB_2035023), COX2 (#66351-1-Ig, Proteintech, RRID:AB_2881731), MMP3 (#14351, Cell Signaling Technology, RRID:AB_2798459), Collagen II (#BA0533, Boster), FTH1 (#4393, Cell Signaling Technology, RRID:AB_11217441), MMP13 (#ab219620, Abcam), TRPV1 (#66983-1-Ig, Proteintech, RRID:AB_2882302), GPX4 (#ab125066, Abcam, RRID:AB_10973901) and GAPDH (#5174, Cell Signaling Technology, RRID:AB_10622025) (Supplementary Table 4). After horseradish peroxidase-conjugated goat anti-rabbit/mouse secondary antibodies (#BL003A or #BL001A, Biosharp) incubation, western blots were imaged by the ChemiDocXRS + Imaging System (Tanon, Shanghai, China). Image J (version 1.8.0) was used for the quantitative analysis of proteins. All antibodies used in this study were commercial antibodies.
Lv Z., Han J., Li J., Guo H., Fei Y., Sun Z., Dong J., Wang M., Fan C., Li W., Xie Y., Sun W., Chen J., Liu Y., Chen F., Liu Z., Liu A., Wu R., Xu X., Yan W., Jiang Q., Ikegawa S., Chen X, & Shi D. (2022). Single cell RNA-seq analysis identifies ferroptotic chondrocyte cluster and reveals TRPV1 as an anti-ferroptotic target in osteoarthritis. eBioMedicine, 84, 104258.
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2

Cell Signaling Pathway Inhibitors Assay

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Anisomycin, SB203580 [45 (link)], SP600125, and ferrostatin-1 [46 (link)] were purchased from MCE (Shanghai, China) and dissolved in dimethyl sulfoxide (DMSO). Cells were treated with anisomycin at the indicated concentration or coincubated with SB203580 (20 μM) or ferrostatin-1 (4 μM). Commercially available antibodies were used in this study. The anti-tubulin antibody was obtained from InTech (Shanghai, China), while anti-c-Myc (#18583), CD133 (#64326), Nanog (#4903), EpCAM (#2929), caspase-3 (#14220), Bcl-2 (#15071), Bax (#5023), N-cadherin (#13116), E-cadherin (#3195), vimentin (#5741), α-smooth muscle actin (α-SMA, #19245), p38 MAPK (#8690), phospho-p38 MAPK (p-p38 MAPK, #9216), histone H3 (#4499), phospho-histone H3 (Ser10) (p-H3S10, #53348), phospho-histone H3 (Ser28) (p-H3S28, #9713), NRF2 (#12721), SLC7A11 (#12691), FTH1 (#4393), and NCOA4 (#66849) antibodies were purchased from Cell Signaling Technology (Boston, USA). Anti-CD24 (ab31622) antibody was provided by Abcam (Cambridge, UK).
Chen W., Yang W., Zhang C., Liu T., Zhu J., Wang H., Li T., Jin A., Ding L., Xian J., Tian T., Pan B., Guo W, & Wang B. (2022). Modulation of the p38 MAPK Pathway by Anisomycin Promotes Ferroptosis of Hepatocellular Carcinoma through Phosphorylation of H3S10. Oxidative Medicine and Cellular Longevity, 2022, 6986445.
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3

Investigating Lung Cancer Cell Lines

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The human lung cancer cell lines A549, H157, HCC827, and PC9 were obtained from the American Type Culture Collection and were genotyped and authenticated before experiments. Cells were cultured in RPMI-1640 medium (HyClone) supplemented with 10% fetal bovine serum (ZETA) at 37°C in a humidified incubator with 5% CO2. Purified MA (>98%) (#sc-200733) was purchased from Santa Cruz. The 20 mM stock solution was made in DMSO. N-acetyl-l-cysteine (NAC) (#HY-B0215), necrostatin-1 (#HY-15760), Z-VAD-FMK (#HY-16658B), liproxstatin-1 (#HY-12726), erastin (#HY-15763) and osimertinib (#HY-15772) were purchased from MCE. The antibodies used were as follows: p-ERK (#4370), ERK (#4695), p-AMPK (#2535), AMPK (#5832), GPX4 (#52455), SLC7A11 (#12691), NRF2 (#12721), NCOA4 (#66849), FTH1 (#4393), PERK (#5683), IRE1a (#3294), LC3A/B (#12741) and GRP78 (#3177) were purchased from Cell Signaling Technology. BCL2 (#12789-1-AP), KRAS (#12063-1-AP) and GAPDH (#10494-1-AP) were purchased from Proteintech. PLA2G4A (#sc-454) was purchased from Santa Cruz.
Ni Y., Liu J., Zeng L., Yang Y., Liu L., Yao M., Chai L., Zhang L., Li Y., Zhang L, & Li W. (2023). Natural product manoalide promotes EGFR-TKI sensitivity of lung cancer cells by KRAS-ERK pathway and mitochondrial Ca2+ overload-induced ferroptosis. Frontiers in Pharmacology, 13, 1109822.
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4

Immunofluorescence Analysis of Knee Cartilage

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Mouse knee sections were dewaxed and hydrated in xylene and graded alcohol, and antigen repair was performed using pepsin for 1 h at 37 °C. After washed by phosphate‐buffered saline (PBS), the slides were blocked with 5% bovine serum albumin (BSA) for 1 h at 37 °C. At last, the slices were incubated overnight (4 °C) with primary antibodies (1:200) of p‐Camk II (#12 716, Cell Signaling Technology), Mmp13 (#GB11247, Servicebio), Gpx4 (#A11243, ABclonal), Cox2 (#12 282, Cell Signaling Technology), and Ncoa4 (#66 849, Cell Signaling Technology). After three washes with TBST, the slices were incubated with fluorescein isothiocyanate (FITC)‐coupled secondary antibody for 1 h at 37 °C. After staining the nucleus by 2‐(4‐Amidinophenyl)−6‐indolecarbamidine dihydrochloride (DAPI), the IF images were acquired using a fluorescence microscope (Zeiss, Germany).
Li W., Lv Z., Wang P., Xie Y., Sun W., Guo H., Jin X., Liu Y., Jiang R., Fei Y., Tan G., Jiang H., Wang X., Liu Z., Wang Z., Xu N., Gong W., Wu R, & Shi D. (2024). Near Infrared Responsive Gold Nanorods Attenuate Osteoarthritis Progression by Targeting TRPV1. Advanced Science, 11(16), 2307683.
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