Anti errα

Manufactured by Cell Signaling Technology

Sourced in United Kingdom

Anti-ERRα is a primary antibody that specifically recognizes the Estrogen-Related Receptor alpha (ERRα) protein. ERRα is a nuclear receptor that plays a role in regulating gene expression related to cellular energy metabolism. The Anti-ERRα antibody can be used to detect and quantify ERRα in various experimental applications.

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Lab products found in correlation

Bca kit, by Thermo Fisher Scientific (3 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions) Erlotinib, by Cell Signaling Technology (1 mentions) Anti flag antibody, by Merck Group (1 mentions) Pcdna4 myc pgc 1α, by Addgene (1 mentions) Anti p erk erk, by Cell Signaling Technology (1 mentions) Grem1 antibody, by Abcam (1 mentions) Xct790, by Bio-Techne (1 mentions) Egfr wt, by Addgene (1 mentions) Recombinant human grem1, by R&D Systems (1 mentions) Anti p akt akt, by Cell Signaling Technology (1 mentions) Recombinant human egf, by Merck Group (1 mentions) Ly294002, by Bio-Techne (1 mentions) U0126, by Bio-Techne (1 mentions) Mcf 7, by American Type Culture Collection (1 mentions) Mcf 10a, by American Type Culture Collection (1 mentions) Mda mb 468, by American Type Culture Collection (1 mentions) Skbr3, by American Type Culture Collection (1 mentions) Mda mb 453, by American Type Culture Collection (1 mentions) Anti α tubulin, by Santa Cruz Biotechnology (1 mentions)

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Anti-ERRα antibody (4 citations)

5 protocols using anti errα

GREM1 Regulates EGFR Signaling in Breast Cancer Cell Lines

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MCF-10A, MCF-10A-ras, MDA-MB-453, MDA-MB-468, SKBR3, MCF-7, T47D, and CCD-1068sk cells were originally obtained from American Type Culture Collection, and the BT474 cell line was obtained from Korean Cell Line Bank. The cells were cultured according to the standard procedure and maintained at 37 °C in a humidified atmosphere composed of 5% CO₂/95% air. GREM1 antibody was purchased from Abcam and recombinant human GREM1 was obtained from R&D systems. Recombinant human EGF, anti-Flag antibody and cell linker kits (PKH26 and PKH67) were purchased from Sigma-Aldrich. Anti-ERRα, anti-p-EGFR/EGFR, anti-p-Akt/Akt, anti-p-ERK/ERK antibodies and erlotinib were obtained from Cell Signalling Technology. Expression plasmids of GREM1, ERRα, Flag-only, Flag-EGFR, and Flag-BMP2 were purchased from Sino Biological Inc. Fc-IgG1 and Fc-GREM1 were provided by ACROBiosystems. The lentiviral GREM1 clone was obtained from Genecopoeia. 3xERRE-luciferase, pcDNA4-myc-PGC-1α, EGFR-WT, EGFR-ECD and EGFR-ICD were provided by Addgene. XCT790, LY294002 and U0126 were purchased from Tocris.

Park S.A., Sung N.J., Choi B.J., Kim W., Kim S.H, & Surh Y.J. (2020). Gremlin-1 augments the oestrogen-related receptor α signalling through EGFR activation: implications for the progression of breast cancer. British Journal of Cancer, 123(6), 988-999.

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Immunofluorescence Analysis of FABP5 and ERRα

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Cells transfected with vector, pCI-neo/FABP5 or pCI-neo/NES-FABP5 were fixed with 4% PFA for 20 min (Nacalai Tesque). Cells were incubated with the following primary antibodies overnight at 4°C: anti-FABP5 (1:100) (Cell Signaling Technology), anti-α-tubulin (1:1000) (Santa Cruz Biotechnology), and anti-ERRα (1:100) (Cell Signaling Technology). After washing with TBS twice, cells were incubated with the following secondary antibodies overnight at 4°C: Alexa Fluor^® 488-conjugated donkey anti-rabbit IgG H&G (1:2000) (Abcam) and Alexa Fluor^® 647-conjugated goat anti-mouse IgG H&G (1:2000) (Abcam). Cells were stained with Hoechst 33342 (Nacalai Tesque). Laser scanning confocal microscopy was performed on an Olympus FV1000.

Senga S., Kawaguchi K., Kobayashi N., Ando A, & Fujii H. (2018). A novel fatty acid-binding protein 5-estrogen-related receptor α signaling pathway promotes cell growth and energy metabolism in prostate cancer cells. Oncotarget, 9(60), 31753-31770.

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Western Blot Analysis of Protein Expression

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Cells were lysed using Triton-X lysis buffer, and then the protein concentration was measured by the BCA kit (Thermo Fisher Scientific, USA). 30 μg of protein samples per lane was separated by polyacrylamide gel electrophoresis and then transferred onto PVDF membrane (Thermo Fisher Scientific, USA). 5% BSA (Thermo Fisher Scientific, USA) dissolved in TBST (Beijing Solarbio Science and Technology, China) was used to block the membrane. The band was incubated with the primary antibody overnight at 4°C. Primary antibodies were applied as follows: anti-GPER (1:5000 dilution, Abcam, UK), anti-ERRα (1:5000 dilution, Cell Signaling Technology, USA), anti-GAPDH (1:5000 dilution, Cell Signaling Technology, USA), and anti-β-actin (1:5000 dilution, Cell Signaling Technology). Secondary antibody (1:5000 dilution, Santa Cruz Biotechnology) incubation was carried out at room temperature for 2h. Protein bands were then detected using chemiluminescence reagent (Millipore, UK) on the ChemiDocTM imaging system (Biorad, USA) and analyzed semi-quantitatively using the Image lab software (Biorad, USA). β-actin and GAPDH were served as internal control.

Ye S., Xu Y., Wang L., Zhou K., He J., Lu J., Huang Q., Sun P, & Wang T. (2020). Estrogen-Related Receptor α (ERRα) and G Protein-Coupled Estrogen Receptor (GPER) Synergistically Indicate Poor Prognosis in Patients with Triple-Negative Breast Cancer. OncoTargets and therapy, 13, 8887-8899.

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Protein Expression Analysis by Western Blot

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Protein was extracted from cells cultured in 6-well plates using RIPA lysis buffer (Thermo Fisher Scientific, 89900) with protease inhibitors (Thermo Fisher Scientific, 78440). A total of 30-μg protein lysates was separated by 4%–12% NuPage Bis-Tris gels (Thermo Fisher Scientific, NP0336) and then transferred to polyvinylidene fluoride membranes (Bio-Rad, 1620264). The membranes were blocked with 5% BSA (Sigma-Aldrich, A7906–100G), probed with primary antibodies overnight at 4°C, and then incubated with the relevant secondary antibody for 1 hour at room temperature. Bands were visualized using ECL (Thermo Fisher Scientific, 34080) with Bio-Rad imaging system. The antibodies of anti-PARP (9532), anti-Rb (9309), anti-pRb^Ser807/811 (8516), anti-γH2AX (9718), anti-TFAM (8076), anti-AMPKα (5831), anti-pAMPKα^Thr172 (2535), anti-ERRα (13826), anti-VDAC (4661), anti-PHB1 (2426), anti-HSP60 (12165), anti-NRF2 (12721), anti-SDHA (11998), anti–β-actin (12620), anti–rabbit-IgG-HRP (7074), and anti–mouse-IgG-HRP (7076) were purchased from Cell Signaling Technology. The antibodies of anti-PGC1α (66369), anti-UQCRC1 (21705), anti-ATP5A1 (14676), anti-COXIV (11242), anti-NDUFB8 (14794), and anti-TUFM (26730) were purchased from Proteintech. Two or three independent experiments were performed.

Wei S., Yin D., Yu S., Lin X., Savani M.R., Du K., Ku Y., Wu D., Li S., Liu H., Tian M., Chen Y., Bowie M., Hariharan S., Waitkus M., Keir S.T., Sugarman E.T., Deek R.A., Labrie M., Khasraw M., Lu Y., Mills G.B., Herlyn M., Wu K., Liu L., Wei Z., Flaherty K.T., Abdullah K., Zhang G, & Ashley D.M. (2022). Antitumor Activity of a Mitochondrial-Targeted HSP90 Inhibitor in Gliomas. Clinical cancer research : an official journal of the American Association for Cancer Research, 28(10), 2180-2195.

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Western Blot Analysis of Mitochondrial Proteins

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Brain tissues and cultured cells were harvested and then lysed with RIPA buffer (Solarbio, China) containing cOmplete™ protease inhibitor cocktail (Roche). Proteins were electrophoretically separated on 10% SDS-PAGE gels and then transferred onto PVDF membranes (Millipore, USA). After blocking with 5% nonfat milk, the membranes were incubated with primary antibodies overnight at 4 °C. The primary antibodies included anti-PGC-1α (1:1000, Thermo Fisher), anti-LC3 (1:1000, Cell Signaling Technology), anti-ERRα (1:1000, Cell Signaling Technology), anti-ULK1 (1:1000, Cell Signaling Technology), anti-sequestosome1 (SQSTM1) (1:1000, Cell Signaling Technology), anti-TOMM20 (1:1000, Cell Signaling Technology), anti-COX IV (1:1000, Abcam), anti-IMMT (1:1000, Abcam), anti-NLRP3 (1:1000, AdipoGen), anti-ASC (1:200, Santa Cruz), and anti-IL-1β (1:800, AdipoGen). After washing, the specific blots were incubated with the species-appropriate secondary antibodies for 1 h at room temperature. Finally, the protein bands were viewed with a Gel Doc 2000 imaging system (Bio-Rad, USA) and then analyzed with the ImageJ software. In the quantitative analysis of Western blots, all the bands detected were within the linear range of detection.

Han B., Jiang W., Cui P., Zheng K., Dang C., Wang J., Li H., Chen L., Zhang R., Wang Q.M., Ju Z, & Hao J. (2021). Microglial PGC-1α protects against ischemic brain injury by suppressing neuroinflammation. Genome Medicine, 13, 47.

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