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Ab213556

Manufactured by Abcam
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Ab213556 is a laboratory equipment product. It serves as a core function for specific laboratory applications.

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Lab products found in correlation

Ab192890, by Abcam (2 mentions) Ab109272, by Abcam (1 mentions) Ab109268, by Abcam (1 mentions) Doxorubicin, by Merck Group (1 mentions) Anti phospho akt ser473, by Cell Signaling Technology (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions) Anti parkin, by Cell Signaling Technology (1 mentions) 3 methyladenine 3 ma, by MedChemExpress (1 mentions) Anti nanog, by Cell Signaling Technology (1 mentions) Anti optineurin, by Abcam (1 mentions) Ab12337, by Abcam (1 mentions) Anti p mtor antibody, by Abcam (1 mentions) Anti sqstm1 p62, by Cell Signaling Technology (1 mentions) Cyclosporin a, by MedChemExpress (1 mentions) Anti lc3 1 2, by Cell Signaling Technology (1 mentions) Anti ki67, by Cell Signaling Technology (1 mentions) Anti β actin, by Cell Signaling Technology (1 mentions) Bortezomib, by Selleck Chemicals (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) Ab32127, by Abcam (1 mentions)

5 protocols using ab213556

1

Immunoblotting Analysis of Autophagy Markers

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The following primary antibodies were used in this study: anti-PIWIL1 antibody (ab12337; Abcam, Cambridge, UK), anti-p-mTOR antibody (ab109268; Abcam), anti-LC3 I/II (#2775S; Cell Signaling Technology, Danvers, MA, USA), anti-P62/SQSTM1 (#8025S; Cell Signaling Technology), anti-Parkin (#4211; Cell Signaling Technology), anti-phosphor-NAK/TBK (ab109272; Abcam), anti-NAK/TBK (A5497; Bimake), anti-optineurin (ab213556; Abcam), anti-β-actin (#4970; Cell Signaling Technology), anti-Akt (#4691; Cell Signaling Technology), anti-phospho-Akt (Ser473) (#9271; Cell Signaling Technology), anti-Ki67 (#4685; Cell Signaling Technology), anti-Bcl-2 (A5010; Bimake), anti-OCT4 (60242-1-Ig; Proteintech), and anti-Nanog (#4903T; Cell Signaling Technology). Goat anti-mouse and goat anti-rabbit antibodies (Biosciences) were used as the secondary antibodies for western blotting. Dexamethasone and doxorubicin were purchased from Sigma (St. Louis, MO, USA), and bortezomib was purchased from Selleck Chemicals. 3-Methyladenine (3-MA) and the mitophagy inhibitor cyclosporin A (CsA) were purchased from MedChemExpress (NJ, USA). All were dissolved in dimethyl sulfoxide (DMSO) and stored at −20°C for up to 6 months. For all cell-based experiments, drugs were diluted at least by 1:1000 to ensure that the final DMSO concentration was lower than 0.1%.
Wang Y., Yao L., Teng Y., Yin H, & Wu Q. (2022). PIWIL1 Drives Chemoresistance in Multiple Myeloma by Modulating Mitophagy and the Myeloma Stem Cell Population. Frontiers in Oncology, 11, 783583.
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2

Protein Expression Analysis in SK-N-SH Cells

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The total protein was extracted from SK-N-SH cells with RIPA containing
phenylmethanesulfonyl fuoride (PMSF) and proteinase inhibitors. Total protein
concentration was measured with the BCA protein assay kit. Equal amounts of
protein (20 μg) were separated by sodium dodecyl sulfate/polyacrylamide gel
electrophoresis and transferred to a polyvinylidene fluoride membrane. The
membrane was blocked in 5% nonfat dry milk for 2 h and probed with primary
antibodies against PINK1 (CST6946, 1:1000, Cell Signal Technology, Boston, MA),
Parkin (CST4211, 1:1000), Optineurin (ab213556, 1:1000, Abcam, Waltham, MA), LC3
(ab192890, 1:2000, Abcam), α-Synuclein (ab32127, 1:20000, Abcam), overnight at
4°C. After washing with TBST, the membranes were incubated for 1 h with a
horseradish-peroxidase-conjugated anti-rabbit/mouse IgG (GB-2301/GB-2305,
1:10000, ZSGB-BIO, Beijing, China). Protein bands were visualized using the
enhanced ECL reagent and quantified with Quantity One.
Zhang Y., Hu H.T., Cao Y.M., Jiang Z.G., Liu J, & Fan Q.Y. (2023). Biphasic Dose-Response of Mn-Induced Mitochondrial Damage, PINK1/Parkin Expression, and Mitophagy in SK-N-SH Cells. Dose-Response, 21(2), 15593258231169392.
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3

Plasma Aβ40/42 and Platelet OPTN Levels

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Plasma Aβ40 and Aβ42 were detected by a commercial ELISA kit (E‐EL‐H0542c 90 and E‐EL‐H0543c, Elabscience, China). The platelet OPTN level was detected by Western blotting (ab213556, Abcam) routinely used in our laboratory.
Yu H., Liu Y., He T., Zhang Y., He J., Li M., Jiang B., Gao Y., Chen C., Ke D., Liu J., He B., Yang X, & Wang J. (2021). Platelet biomarkers identifying mild cognitive impairment in type 2 diabetes patients. Aging Cell, 20(10), e13469.
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4

Protein Extraction and Western Blot Analysis

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Total protein from cells or brain tissues was isolated by RIPA buffer (Beyotime). In addition, mitochondria were isolated from the cytoplasm using the Cell Mitochondria Isolation Kit (Beyotime) and incubated with the mitochondrial lysis buffer supplemented with PMSF (Beyotime) to extract mitochondrial proteins. After quantification using BCA Protein Quantification Kit (Thermo Fisher Scientific), equal amounts of proteins were loaded onto an SDS-PAGE gel for separation. Subsequently, the protein signal was transferred onto the PVDF membrane and blocked by 5% non-fat milk powder. The membrane was incubated with primary antibodies against SENP1 (1/1000; AF0275, Affinity), NeuN (1/2000; ab177487, Abcam), GFAP (1/10000; ab7260, Abcam), LC3 (1/2000; ab192890, Abcam), p62 (1/10000, ab109012, Abcam), OPTN (1/1000; ab213556, Abcam), COX IV (1/2000; ab202554, Abcam) and GAPDH (1/10000; ab181602, Abcam) overnight at 4°C. After incubating with appropriate secondary antibody at 37°C for 1 h, the blot was visualized using ECL detection system (Thermo Fisher Scientific). Relative protein levels were assessed by ImageJ software.
He J., Peng J., Li Y., Jiang J., Li J., Lin L., Wang J, & Xia Y. (2024). SENP1 facilitates OM-MSC differentiation through activating OPTN-mediated mitophagy to mitigate the neurologic impairment following ICH. iScience, 27(6), 109865.
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5

Endogenous OPTN SUMOylation Assay

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To measure the SUMOylation of OPTN in endogenous level, transfected OM-MSCs were washed three times with cold PBS and then cleaved in IP lysis buffer. Cell lysates were centrifuged (12000 g, 4°C) and supernatants were precipitated by OPTN antibody (1/40; ab213556, Abcam) and IgG (negative control). Protein A/G agarose beads were added to the cells and incubated overnight at 4°C. The washed precipitated proteins were detected by Western blotting with SUMO1 (1/1000; ab32058, Abcam), SENP1 and OPTN.
He J., Peng J., Li Y., Jiang J., Li J., Lin L., Wang J, & Xia Y. (2024). SENP1 facilitates OM-MSC differentiation through activating OPTN-mediated mitophagy to mitigate the neurologic impairment following ICH. iScience, 27(6), 109865.
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