Anti hmgcr

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-HMGCR is a laboratory product that detects the presence of the HMGCR (3-hydroxy-3-methylglutaryl-CoA reductase) protein. HMGCR is an enzyme involved in the production of cholesterol. This product can be used to measure HMGCR levels in various biological samples.

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Close spelling variants of this product

HMGCR (8 citations) Anti-HMGCR antibody (3 citations) Anti-HMGCR (sc-271595) (2 citations)

10 protocols using anti hmgcr

Western Blot Analysis of Metabolic Regulators

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Mouse liver, skeletal muscle or white adipose tissue were dissected and immediately frozen in liquid nitrogen. Whole-cell extracts were prepared using lysis buffer (10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA, 1% Igepal CA-630, 0.2mM Na₃VO₄ and a protease inhibitor cocktail (Roche Diagnostics)). Protein concentration was measured by colorimetric assay (Bio-Rad Laboratories, Richmond, CA, USA) and equal amount of proteins was loaded onto SDS gels. After transfer to polyvinylidene difluoride membranes, proteins were probed with primary antibodies (1 μg/ml), followed by horseradish peroxidase-conjugated secondary antibodies, washed and visualized with SuperSignal West Pico/Dura chemiluminescent substrate (Pierce-Thermo Fisher Scientific, Waltham, MA, USA). Blots were reprobed with β-actin-specific antibody for loading controls.
Anti-IRS-1 (Santa Cruz Biotechnology), anti-IRS-2 (Santa Cruz Biotechnology), anti-SCD-1 (Santa Cruz Biotechnology), anti-IRβ (Santa Cruz Biotechnology), anti-pAkt2 (Cell Signaling Technology, Danvers, MA, USA), anti-SREBP-1 (Santa Cruz Biotechnology), anti-ACCα (Santa Cruz Biotechnology), anti-FAS (Santa Cruz Biotechnology), anti-SREBP-2 (Santa Cruz Biotechnology), anti-HMGCR (Santa Cruz Biotechnology), anti-Albumin (Santa Cruz Biotechnology) and anti-β-actin (Santa Cruz Biotechnology) antibodies were used as primary antibodies.

Huo J., Ma Y., Liu J.J., Ho Y.S., Liu S., Soh L.Y., Chen S., Xu S., Han W., Hong A., Lim S.C, & Lam K.P. (2016). Loss of Fas apoptosis inhibitory molecule leads to spontaneous obesity and hepatosteatosis. Cell Death & Disease, 7(2), e2091-.

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Western Blot Analysis of Signaling Proteins

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Primary antibodies used in this study were purchased from Cell Signaling Technology (Beverly, MA): anti-pPKA^Thr198/PKA, anti-pCREB^Ser133/CREB, anti-pSTAT3^Tyr705/STAT3, anti-pSrc^Ser17/Src, anti-pACC ^Ser79/ACC, anti-pAkt^Ser473/Akt, anti-pAMPK ^Thr172/AMPK, anti-p4EBP1^Thr37/46/4EBP1, anti-pmTOR^Ser2448/mTOR, anti-pP70S6K^Thr389/P70S6K, anti-EPAC-1, anti-BEATA2_AR, anti-FAK, anti-FASN, anti-GLUT4, anti-OCT3, anti-NOTCH, anti-PGC1, anti-pPRAS40^Thr246, anti-PRAS40, anti-pRAPTOR^Ser792, anti-RAPTOR, and anti-PI3K110α. Anti-BCL-2 was purchased from BD Bioscience (San Jose, CA). Anti-P27 was purchased from Thermo Fisher Scientific (Waltham, MA). Anti-rabbit immunoglobulin-horseradish peroxidase-conjugated secondary antibody, LumiGLO reagent with peroxide and cAMP assay kit were also purchased from Cell Signaling Technology, Inc. (Beverly, MA). Anti-IGF1Rα, anti-HMGCR, anti-P21, anti-SCD1, and anti-SCEBP1 were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX); mouse anti-β-actin primary antibody was obtained from Sigma Aldrich (St. Louis, MO). The 1-methyl-1-nitrosourea (MNU) was obtained from Ash Stevens (Detroit, MI) and stored at −80°C prior to use. Metformin and buformin were obtained from Waco Pure Chemical Industries, (Waco, TX); phenformin was obtained from Sigma Aldrich (St. Louis, MO).

Zhu Z., Jiang W., Thompson M.D., Echeverria D., McGinley J.N, & Thompson H.J. (2015). Effects of Metformin, Buformin, and Phenformin on the Post Initiation Stage of Chemically-Induced Mammary Carcinogenesis in the Rat. Cancer prevention research (Philadelphia, Pa.), 8(6), 518-527.

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Immunohistochemical Analysis of Metastatic Lung Tissues

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IHC staining was performed on formalin-fixed and paraffin-embedded sections of lung metastases with antibodies of anti-HMGCR (1:100, Santa cruz), anti-CCDC25 (1:100, Cell Signaling Technology), anti-MPO (1:200, R&D), anti-H3cit (1:300, Cell Signaling Technology), or anti-Caveolin-1 antibody (1:100, Cell Signaling Technology). Details can be found in the supplementary data.
Freshly obtained mouse lung tissues were quick-frozen in liquid nitrogen and placed in a cryostat to prepare frozen sections, each with a thickness of 4 mm. For IF staining, slides were rewarm at room temperature for 30 min and then combined with antibodies against CCDC25, Caveolin-1, MPO and H3cit overnight at 4 °C. Then rinsed the sections with ice PBS, and combined with secondary antibodies, respectively. Added DAPI-containing mounting medium, followed by mounting with a coverslip.

Tang Q., Liang B., Zhang L., Li X., Li H., Jing W., Jiang Y., Zhou F., Zhang J., Meng Y., Yang X., Yang H., Huang G, & Zhao J. (2022). Enhanced CHOLESTEROL biosynthesis promotes breast cancer metastasis via modulating CCDC25 expression and neutrophil extracellular traps formation. Scientific Reports, 12, 17350.

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Immunoblotting Protein Analysis in Cells

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Immunobloting analysis of proteins in cell lysates and nuclear fractions was performed as previously described [56 (link), 57 (link)]. Primary antibodies used were as follows: anti-YAP (#4912), anti-p-YAP (#4911), anti-Myc-Tag (#2278), anti-Mst (#3682), anti-p-Mst (#3681), anti-LATS (#3477) and anti-p-LATS (#8654) were purchased from Cell Signaling Technology (Beverly, MA); anti-HIF-1α (#610959) was purchased from BD; anti-β-Actin (#Sc-47778), anti-Lamin B (#Sc-6216), anti-BNIP3 (#Sc-56167), and anti-HMGCR (#27578) were purchased from Santa Cruz (Santa Cruz, CA).

Dai X.Y., Zhuang L.H., Wang D.D., Zhou T.Y., Chang L.L., Gai R.H., Zhu D.F., Yang B., Zhu H, & He Q.J. (2016). Nuclear translocation and activation of YAP by hypoxia contributes to the chemoresistance of SN38 in hepatocellular carcinoma cells. Oncotarget, 7(6), 6933-6947.

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Antibody-based Analysis of Lipid Metabolism

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The following antibodies were used: guinea pig and mouse monoclonal against Adipose Differentiation Related Protein (ADRP; Research Diagnostics, Concord, MA, USA), monoclonal anti-α-tubulin (BD Transduction Laboratories, Franklin Lakes, NJ, USA), CS-35 anti-lipoarabinomannan (LAM) monoclonal, rabbit anti-whole ML (kindly provided by Dr Patrick J. Brennan, Colorado State University, Fort Collins, CO, USA; NIH/NIAID contract 1Al25469), anti-SR-A1 (mouse monoclonal, SC-166184), SREBP-1 (rabbit polyclonal, SC-8984), LDL-R (goat polyclonal, SC-11824), anti-HMGCR (SC-33827, rabbit polyclonal IgG) and anti-GADPH (mouse monoclonal) from Santa Cruz Biotechnology (Santa Cruz, CA, USA), anti-CD36 (rabbit polyclonal; Cayman Chemical, Ann Arbor, MI, USA) and anti-CD36 (rat monoclonal, Ab80080), SREBP-2 (rabbit polyclonal, ab30682) from Abcam (Cambridge, MA, USA), fluorescent-labelled (Alexa Fluor 488, 555, and 633) goat anti-rabbit and anti-mouse (Molecular Probes, Eugene, OR, USA), donkey anti-guinea pig fluorescent-dye-Cy2 conjugated, and, lastly, control IgG (Jackson ImmunoResearch Laboratories, West Grove, PA, USA).

Mattos K.A., Oliveira V.C., Berrêdo-Pinho M., Amaral J.J., Antunes L.C., Melo R.C., Acosta C.C., Moura D.F., Olmo R., Han J., Rosa P.S., Almeida P.E., Finlay B.B., Borchers C.H., Sarno E.N., Bozza P.T., Atella G.C, & Pessolani M.C. (2014). Mycobacterium leprae intracellular survival relies on cholesterol accumulation in infected macrophages: a potential target for new drugs for leprosy treatment. Cellular Microbiology, 16(6), 797-815.

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Antibody Panel for Cell Signaling

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The anti-FLAG and anti-caspase-11 antibodies were from Sigma. The anti-SREBP1, anti-HMGCS1, anti-HMGCR antibodies were from Santa Cruz Biotechnology. The anti-HA antibody was from Roche, and the anti-S1P and anti-GAPDH antibodies were from Abcam. The anti-RCAS1, anti-EEA1, anti-LAMP1, anti-human caspase-4 and anti-caspase-5 antibodies were from Cell Signaling Technology. The LPS was purchased from Sigma (L2637).

Cheng Y., Manabe I., Hayakawa S., Endo Y, & Oishi Y. (2023). Caspase-11 contributes to site-1 protease cleavage and SREBP1 activation in the inflammatory response of macrophages. Frontiers in Immunology, 14, 1009973.

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Western Blotting Analysis of ABC Transporters

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Whole-cell lysates were prepared with RIPA Lysis Buffer (Beyotime, P0013B, China), then the proteins were separated using 10% SDS-polyacrylamide gels, and to 0.45 μm PVDF membranes, then blocked and incubated with the following primary antibodies: Anti-SIAH1 (Abcam, ab69638, 1:200), anti-GAPDH (Cell Signaling, D16H11, 1:1000). Anti-ABCB1 (sc-55,510, 1:1000), anti-ABCB4 (sc-58,221, 1:1000), anti-HMGCR (sc-271,595, 1:1000) were purchased from Santa Cruz Biotechnology; anti-ABCG1 (13578-1-AP, 1:1000), anti-ABCG2 (27286-1-AP, 1/1000 dilution) were from Proteintech Group. Then membranes were incubated with secondary antibodies (Cell Signaling Technology) and incubated with the ECL western blotting substrate. Chemiluminescent images were acquired with the iBright CL1000.

Yuan H., Wu H., Cheng J, & Xiong J. (2023). SIAH1 ubiquitination-modified HMGCR inhibits lung cancer progression and promotes drug sensitivity through cholesterol synthesis. Cancer Cell International, 23, 71.

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Gp78 Antibody Generation and Characterization

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Gp78 mouse monoclonal antibody was purchased from Abnova. Polyclonal anti-gp78 antibodies were generated in immunized rabbit with recombinant protein of gp78 cytosolic domain (309–643 aa) and peptide (524–537 aa), and purified with affinity chromatography. Anti-insig-1/-2, anti-SREBP-1, anti-HMGCR, and anti-fatty acid synthase, anti-GRP78 antibodies were obtained from Santa Cruz. Anti-PDI and anti-Chop were obtained from Cell Signaling Technology. Anti-actin and Oil-red O was purchased from Sigma. Masson’s Trichrome Stain Kit was purchased from Polysciences.

Zhang T., Kho D.H., Wang Y., Harazono Y., Nakajima K., Xie Y, & Raz A. (2015). Gp78, an E3 Ubiquitin Ligase Acts as a Gatekeeper Suppressing Nonalcoholic Steatohepatitis (NASH) and Liver Cancer. PLoS ONE, 10(3), e0118448.

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Western Blot Analysis of Protein Signaling

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HepG2 cells were lysed in ice with Complete Tablet Buffer (Roche, Basel, Switzerland) supplemented with 2 mM sodium orthovanadate (Na3VO4), 1 mM phenylmethanesulfonyl fluoride (PMSF) (Sigma-Aldrich, St. Louis, MO, USA), 1:50 mix Phosphatase Inhibitor Cocktail (Sigma-Aldrich, St. Louis, MO, USA), and 1:200 mix Protease Inhibitor Cocktail (Sigma-Aldrich, St. Louis, MO, USA). According to the standard protocol, 35 μg of protein of each sample was resolved on 10% SDS-PAGE gels, and polyvinylidene difluoride membranes (PVDF, GE, Healthcare Europe GmbH) was incubated overnight at 4 °C with the following specific primary antibodies: anti-LDL receptor (1:500; Santa Cruz, CA, USA), anti-pSRC (1:500; Santa Cruz, CA, USA), anti-SRC (1:500; Santa Cruz, CA, USA), anti-pAMPK (1:500; Santa Cruz, CA, USA), anti-AMPK (1:500; Santa Cruz, CA, USA) and anti-HMGCr (1:500; Santa Cruz, CA, USA). Protein expression was normalized and verified through anti-β-actin (Sigma-Aldrich, St. Louis, MO, USA). The results were expressed as means ± SD (% vs. control).

Ruga S., Galla R., Penna C., Molinari C, & Uberti F. (2022). The Activity of Ten Natural Extracts Combined in a Unique Blend to Maintain Cholesterol Homeostasis—In Vitro Model. International Journal of Molecular Sciences, 23(7), 3805.

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Characterizing Lipid Raft Dynamics in Cells

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Alexa Fluor 488-conjugated cholera toxin subunit B (CTxB) was obtained from Molecular Probes (Eugene, OR, USA).
The anti-EGFR, anti-Src, anti-HMG-CR, anti-sterolregulatory element-binding protein (SREBP)-1, and horseradish peroxidase (HRP)-conjugated goat anti-mouse and anti-rabbit antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-phospho-Src and anti-fatty acid synthase (FASN) antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). Antiphospho-EGFR (1068) was obtained from Invitrogen/Life technologies (Carlsbad, CA, USA). Doxorubicin, methyl-βcyclodextrin (MβCD), water-soluble cholesterol, and simvastatin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mevalonolactone, which turns to mevalonate [20] , was purchased from Sigma-Aldrich.

Yun U.J., Lee J.H., Shim J., Yoon K., Goh S.H., Yi E.H., Ye S.K., Lee J.S., Lee H., Park J., Lee I.H, & Kim Y.N. (2019). Anti-cancer effect of doxorubicin is mediated by downregulation of HMG-Co A reductase via inhibition of EGFR/Src pathway. Laboratory investigation; a journal of technical methods and pathology, 99(8).

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