Clusterin

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Clusterin is a laboratory product offered by Santa Cruz Biotechnology. It is a glycoprotein that plays a role in various cellular processes. The core function of Clusterin is to act as a molecular chaperone, assisting in the folding and stabilization of proteins.

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Lab products found in correlation

6 protocols using clusterin

Alisol A 24-acetate Induces Autophagy in Renal Cells

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Trypsin was provided by KeyGEN Biotech Co., Ltd (Nanjing, China). Dulbecco’s modified eagle medium (DMEM)-F12 (1:1) medium with high-glucose and fetal bovine serum (FBS) were purchased from Gibco/BRL (Grand Island, New York, NY, USA). 3-4,5-dimethyl-2-thiazolyl)-2,5- diphenyl-2-H-tetrazolium bromide (MTT), dimethylsulfoxide (DMSO), and 3-methyl adenine (3-MA) were provided by Sigma Chemical (St. Louis, MO, USA). Alisol A 24-acetate (24A) and Alisol B 23-acetate (23B; purity ≥95%) were purchased from Tianjin Evans Science and Technology Co., Ltd (Tianjin, China). Primary and second antibodies against LC3, Beclin-1, Bcl-2, Bcl-xl, Clusterin, Kim-1, TFF-3 and β-actin were provided by Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Hematoxylin-eosin staining solution was offered by Shanghai Yuanmu Biotechnology Co., Ltd (Shanghai, China). EliVision plus and DAB kits were offered by MXB Biological Technology Co., Ltd (Fuzhou, Fujian, China). Monopotassium phosphate, disodium hydrogen phosphate, sodium chloride, and potassium chloride were obtained from Nanjing Nanao Science and Technology Co., Ltd (Nanjing, China).

Wang C., Feng L., Ma L., Chen H., Tan X., Hou X., Song J., Cui L., Liu D., Chen J., Yang N., Wang J., Liu Y., Zhao B., Wang G., Zhou Y, & Jia X. (2017). Alisol A 24-Acetate and Alisol B 23-Acetate Induced Autophagy Mediates Apoptosis and Nephrotoxicity in Human Renal Proximal Tubular Cells. Frontiers in Pharmacology, 8, 172.

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Western Blot Antibody Detection Protocol

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The same method was applied as with TTR for the following antibodies; Clusterin (Santa Cruz Biotechnology anti-rabbit sc-8354 1/500) GRP78 (BiP) (Santa Cruz Biotechnology anti-rabbit sc-13968 1/1000) and ubiquitin (Santa Cruz Biotechnology anti-rabbit sc-9133 1/1500).

Panayiotou E., Papacharalambous R., Antoniou A., Christophides G., Papageorgiou L., Fella E., Malas S, & Kyriakides T. (2016). Genetic background modifies amyloidosis in a mouse model of ATTR neuropathy. Biochemistry and Biophysics Reports, 8, 48-54.

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Gastric Tissue Histology and Immunohistochemistry

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Formalin-fixed paraffin-embedded stomach tissues were processed for histology and immunohistochemistry by standard procedures. Primary antibodies used for immunostaining were: GFP (Invitrogen, A11122, 1:200), Ki67 (Abcam, ab16667, 1:100), Mist1 (Santa Cruz, sc-80984, 1:100), Jagged1 (Santa Cruz, sc-6011, 1:100), Notch1 (Cell Signaling, No. 3608, 1:100), Notch2 (DSHB, University of Iowa, C651.6DbHN, 1:200), Notch3 (ProteinTech, 55114-1-AP, 1:100), Notch4 (Millipore, 09-089, 1:100), Cytokeratin 19 (Abcam, ab52625, 1:200), Muc5AC (Santa Cruz, sc-21701, 1:100), Clusterin (Santa Cruz, sc-6420, 1:100), and TFF3 (ProteinTech, 23277-1-AP, 1:100). X-Gal staining was performed as previously described [18] (link).

Chung W.C., Zhou Y., Atfi A, & Xu K. (2019). Downregulation of Notch Signaling in Kras-Induced Gastric Metaplasia. Neoplasia (New York, N.Y.), 21(8), 810-821.

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Histological Analysis of Mouse Pancreas

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Mouse pancreas tissues were fixed overnight in 4% paraformaldehyde (Sigma-Aldrich, Cat# 158127). Haematoxylin and eosin (H & E) staining and immunostaining were performed to analyse representative cross-sections of the pancreatic head, body and tail. Immunostaining was conducted according to standard protocols [30 (link)]. In brief, endogenous peroxide activity was blocked with 3% peroxidase, and the slides were then incubated with 5% BSA to prevent nonspecific binding. The slides were incubated with primary antibodies at an optimal dilution (clusterin: Santa Cruz Biotechnology, Cat# sc-8354, 1:100; cytokeratin 19: Abcam, Cat# ab15463, 1:100; chymotrypsin: Abcam, Cat# ab35694, 1:100; and amylase: Santa Cruz Biotechnology, Cat# sc-46657, 1:100) and appropriate secondary antibodies. The horseradish peroxidase polymer detection system (PV-9001; GBI Labs, Mukilteo, WA, USA) was employed to detect antibody staining. Five to ten randomly selected 100X H&E images were quantified by a pathologist using standard pathological criteria [31 (link), 32 (link)].

Hong X., Zhang J., Wu Q., Wang W., Ye A.Y., Song W., Dai H., Wang X., Wu F., You L., Wu W, & Zhao Y. (2016). Challenges in detecting pre-malignant pancreatic lesions during acute pancreatitis using a serum microRNA assay: a study based on KrasG12D transgenic mice. Oncotarget, 7(16), 22700-22710.

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Antibody and Reagent Characterization Protocol

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The following antibodies were used: TRPC6 from Alomone (ACC-017)12 (link), Sigma (PRS3889)19 (link) and SAB (21403), TRPC5 from Sigma (ACC-020)48 (link), Ca_v1.2 from Millipore (AB5156), Ca_v3.1 (ACC-021) and Ca_v3.3 (ACC-009) from Alomone, APP from Invitrogen (13-0200, clone LN27) and Sigma (A8717), 6E10 from Chemicon (MAB1560) and Signet (SIG-39300), MOAB-2 from Kerafast (EB2001), ADAM10 from Abcam (Ab1997), clusterin from Santa Cruz (sc-6420), BACE1 from Millipore (MAB5308), insulin-degrading enzyme from EMD Biosciences (PC730), neprilysin from Epitomics (2569–1), PS1 from Chemicon (MAB5232), nicastrin from Sigma (MAB5232), presenilin enhancer 2 from Invitrogen (36–7100), E-Cadherin from Abcam (ab53033), N-Cadherin from BD (610921), cleaved-Notch from Cell Signalling Technology (2421 L), Notch from Proteintech (10062-2-AP), EEA1 from BD (610457), Calnexin from Enzo (ADI-SPA-860-F), GM130 from BD (610823), Bip from BD (610979), Cathepsin D from Santa Cruz (sc-6487), Myc from Chemicon (05–724), HA from Sigma (H6908) and Genscript (A00168), and His from Abmart (M30111). OAG (O6754), SKF96365 (S7809) and L685,458 (L1790) were from Sigma.

Wang J., Lu R., Yang J., Li H., He Z., Jing N., Wang X, & Wang Y. (2015). TRPC6 specifically interacts with APP to inhibit its cleavage by γ-secretase and reduce Aβ production. Nature Communications, 6, 8876.

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Comprehensive Protein Analysis via SDS-PAGE

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Lysed samples were loaded onto two 4 to 20% gradient SDS-PAGE gels (Bio-Rad) and transferred to nitrocellulose membranes (Bio-Rad) by using standard methods. Gels were blocked with 5% bovine serum albumin in PBS with 0.05% Tween 20 (PBST). Antibodies used for Western blotting included 1:500 Tspan8 (sc-292058, Santa Cruz Biotechnology Inc.), 1:1000 TSG101 (ab83, Abcam), 1:1000 CD9 (ab92726, Abcam), 1:1000 GM130 (sc-55591, Santa Cruz Biotechnology Inc.), 1:1000 HSP70 (sc-32239, Santa Cruz Biotechnology Inc.), 1:1000 galectin-3–binding protein (sc-74970, Santa Cruz Biotechnology Inc.), 1:1000 clusterin (sc-8354, Santa Cruz Biotechnology Inc.), 1:1000 filamin-A (sc-28284, Santa Cruz Biotechnology Inc.), 1:1000 glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (2118L, Cell Signaling Technology), 1:1000 Itsn2 (ab133854, Abcam), and 1:1000 CD49d (PA520595, Invitrogen).

Liu Y., Fan J., Xu T., Ahmadinejad N., Hess K., Lin S.H., Zhang J., Liu X., Liu L., Ning B., Liao Z, & Hu T.Y. (2020). Extracellular vesicle tetraspanin-8 level predicts distant metastasis in non–small cell lung cancer after concurrent chemoradiation. Science Advances, 6(11), eaaz6162.

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