Anti atp7a

Manufactured by Santa Cruz Biotechnology

Anti-ATP7A is a laboratory tool used for the detection and analysis of the ATP7A protein, which is involved in copper homeostasis. It provides a means to study the expression, localization, and function of this important cellular component.

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Lab products found in correlation

3 protocols using anti atp7a

Cisplatin Resistance Mechanisms in Cancer Cells

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The sources for various reagents and kits were: Cisplatin (Sigma, St. Louis, MO); hygromycin B (Mediatech, Inc., Herndon, VA); ECL Western blotting detection reagents (GE Healthcare Bio-Sciences Corp., Piscataway, NJ); fetal bovine serum, DMEM, L-glutamine, penicillin, streptomycin, Lipofectamine Plus reagent, Gibco trypsin-EDTA reagent, PCR primers, and nitrocellulose membranes (Invitrogen, Carlsbad, CA); RIPA buffer (Thermo Scientific, Rockford, IL); Mitochondria Isolation Kit and BCA protein assay kit (Pierce, Rockford, IL); Protease inhibitor cocktail (Roche Diagnostics, Indianapolis, IN); TRIzol reagent and methionine-free DMEM (Life Technologies, Grand Island, NY); Wizard Genomic DNA purification kit (Promega, Madison, WI). Sources for antibodies were: anti-Hsp60, (Chemicon International, Temecula, CA); anti-PARP, anti-cleaved caspase 3 and cleaved caspase 7 (Cell Signaling, Beverly, MA); anti-Cisplatin-modified DNA (Abcam, Cambridge, MA); anti-ATP7A, anti-ATP7B, and anti-Ctr1 (Santa Cruz Biotechnology Inc., Santa Cruz, CA). Anti-actin antibody was obtained from Sigma; the antibody recognizes an identical 11-amino acid peptide at the C-terminus of the three major forms, α-, β- and γ-actin. The proteins have identical mobility on SDS gels, and the band is labeled actin in all blots.

Zhang Y, & Maurizi M.R. (2015). Mitochondrial ClpP activity is required for cisplatin resistance in human cells. Biochimica et biophysica acta, 1862(2), 252-264.

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Antibody Characterization for Cell Signaling

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Following antibodies were used at a dilution of 1:1000 except where otherwise stated: Anti-Actin (sc-1616 HRP; 1:10,000), anti-ATP7A (sc-376467; 1:100), anti-normal mouse IgG (sc-2025; 1.4 µg for IP), anti-ERK (sc-514302) from Santa Cruz; anti-GFP (2555), anti-OTULIN (14127; 1.4 µg for IP), anti-IκBα (4814), anti-phospho-IκBα (Ser32/36) (9246), anti-LAMP1 (15665; 1:100), anti-phospho-ERK (Thr202/Tyr204) (9101), anti-normal rabbit IgG (2729; 1.4 µg for IP), anti-EEA1 (3288, 1:100) all CST; anti-SNX27 (ab77799; 1.4 µg for IP and 1:50 for IF), anti-VPS26 (ab23892), anti-VPS35 (ab157220; 1:10,000), anti-MRP4 (ab15602; 1:2500), anti-DGKζ (ab105195; 1:100), anti-GLUT1 (ab15309; 1:100 and ab115730) all Abcam; anti-Beta PIX (07-1450-I; 1:5000), anti-Met1-Ub (MABS199) all Millipore; anti-STEAP3 (17186-1-AP), anti-KIDINS220 (21856-1-AP) all Proteintech; anti-SNX27 (for murine SNX27) gift from W. Hong; anti-Beta-catenin (610153, 1:2000) BD; anti-FLAG M2 (F3165; 1:10000 and 1 µg for IP) Sigma; anti-HOIP (MAB8039; 1:5000) R&D; anti-HA (Core facility monoclonal antibodies HMGU; 25 µl for IP); anti-CD2-APC (17-0029-41; 1:200) eBioscience.

Stangl A., Elliott P.R., Pinto-Fernandez A., Bonham S., Harrison L., Schaub A., Kutzner K., Keusekotten K., Pfluger P.T., El Oualid F., Kessler B.M., Komander D, & Krappmann D. (2019). Regulation of the endosomal SNX27-retromer by OTULIN. Nature Communications, 10, 4320.

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Western Blot Analysis of Protein Extracts

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For protein extraction, cells were lysed in RIPA buffer with protease inhibitor cocktail tablets Complete, EDTA-free (Roche) and phosphatase inhibitor cocktail tablets PhosSTOP (Roche). Lysates were measured for protein concentration by using the Bio-Rad Protein Assay (Bio-Rad), then mixed with Laemmli loading buffer, electrophoresed on SDS-PAGE gels and separated proteins transferred to PVDF blotting membranes (Amersham, GE Healthcare). Membranes were blocked for 1 h in 10% (m/vol) dry milk dissolved in TBS with 1% (vol/vol) Tween-20 (TBSt); incubated with primary antibodies overnight and with secondary antibodies conjugated with horseradish peroxidase, for 1 h. Antibodies were diluted in 10% dry milk in TBSt: anti-HA 1:1000 (Covance), anti-GAPDH 1:40000 (Sigma), anti-p21, anti-p73 1:3000 (Bethyl), anti-ATP7A 1:500 (Santa Cruz). To detect the signal ECL Western Blotting Detection Reagent (Amersham, GE Healthcare) or SuperSignal West Dura Chemiluminescent Substrate (Thermo Scientific) was used.

Lopriore P., Capitanio N., Panatta E., Di Daniele N., Gambacurta A., Melino G, & Amelio I. (2018). TAp73 regulates ATP7A: possible implications for ageing-related diseases. Aging (Albany NY), 10(12), 3745-3760.

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