Anti scd1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-SCD1 is a laboratory reagent that can be used to detect and quantify the presence of the SCD1 (Stearoyl-CoA Desaturase 1) protein in biological samples. SCD1 is an enzyme involved in lipid metabolism. The Anti-SCD1 product can be utilized in various research applications that require the identification or measurement of SCD1 expression levels.

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Goat anti-SCD1 (2 citations) Anti‐SCD1 antibody (2 citations)

8 protocols using anti scd1

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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Gene Expression and Protein Analysis in Biological Research

Cited 1 time

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Total RNA was isolated by using TRIzol reagent (Invitrogen, USA) according to the manufacturer’s instructions. The isolated total RNA was reverse-transcribed by using PrimeScript RT reagent Kit (TaKaRa, Shiga, Japan). Real-time PCR was performed on an ABI 7900 Real-Time PCR System (Applied Biosystems). Specific primers for Real-time PCR were listed in Additional file 5: Table S1. The gene expression is quantified by normalizing to 18s.
Western Blot analysis was performed as described before [40 (link)]. Anti-SCD-1 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-FASN (BD, Biosciences PharMingen, San Diego, CA, USA), anti-ACC1 (Cell Signaling Technology, Danvers, MA), anti-β-Actin (Sigma), anti-α-Tubulin (Sigma), and anti-GYS2 (Cell Signaling Technology) antibodies were used as primary antibodies.

Yao X., Hou S., Zhang D., Xia H., Wang Y.C., Jiang J., Yin H, & Ying H. (2014). Regulation of fatty acid composition and lipid storage by thyroid hormone in mouse liver. Cell & Bioscience, 4, 38.

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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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Western Blot Analysis of Brown Fat Proteins

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Whole-cell extracts were prepared from tissues or brown adipocytes by homogenization in lysis buffer (29 (link)) and subjected to Western blot analysis using the following antibodies: anti–PGC-1α (28 (link)), anti-FAS, anti-SCD1, anti-actin (Santa Cruz, Dallas, TX), anti–α-tubulin, and anti-GAPDH (Abcam, Cambridge, MA).

Jun H.J., Joshi Y., Patil Y., Noland R.C, & Chang J.S. (2014). NT-PGC-1α Activation Attenuates High-Fat Diet–Induced Obesity by Enhancing Brown Fat Thermogenesis and Adipose Tissue Oxidative Metabolism. Diabetes, 63(11), 3615-3625.

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Immunohistochemical Analysis of Lipid Metabolism and Proliferation in Colorectal Cancer

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Human colorectal cancer tissue sections were deparaffinized and rehydrated through graded alcohol. Endogenous peroxidase activity was blocked by incubation in 3% H₂O₂, and antigen retrieval was carried out with 0.01 M citrate buffer (pH 6.0). Immunostaining was performed using the following antibodies: anti-SREBP1 (1:100, Proteintech), anti-SCD1 (1:50, Santa Cruz Biotechnology), and anti-Ki-67 (1:300, Nakasugi Bridge).

Wang H., Chen Y., Liu Y., Li Q., Luo J., Wang L., Chen Y., Sang C., Zhang W., Ge X., Yao Z., Miao L, & Liu X. (2021). The lncRNA ZFAS1 regulates lipogenesis in colorectal cancer by binding polyadenylate-binding protein 2 to stabilize SREBP1 mRNA. Molecular Therapy. Nucleic Acids, 27, 363-374.

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Immunohistochemistry of HBV-infected Liver

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The immunohistochemical analysis of archival paraffin blocks from 5 normal and 155 HBV-infected patients was approved by the Institutional Review Board of the Koo Foundation Sun Yat-Sen Cancer Center, Taipei, Taiwan. The liver tissue arrays from the 155 HBV-infected patients were further stratified by their clinical phases of HBV infection. Tissues samples from mice were collected following the recommendations provided in the Declaration of Helsinki and its amendments.
Human and mouse tissues were formalin fixed, dehydrated, and embedded in paraffin. Deparaffinized sections (4–5 μm) were treated with 1% H₂O₂ to block endogenous peroxidase activity and immersed in boiling 0.01% citric acid (pH 6.0) in a microwave oven for 15 min to enhance antigen retrieval. After cooling, the sections were rehydrated with PBS and stained with hematoxylin and eosin (H&E) or treated with the following antibodies: anti-PML, anti-HBsAg, anti-Ki-67, and anti-Scd1 (all from Santa Cruz Biotechnology, Dallas, Texas, USA). Immunohistochemistry was performed using the ABC Kit (Dako).

Chung Y.L, & Wu M.L. (2018). The Role of Promyelocytic Leukemia Protein in Steatosis-Associated Hepatic Tumors Related to Chronic Hepatitis B virus Infection. Translational Oncology, 11(3), 743-754.

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Protein Extraction and Western Blot Analysis

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To obtain protein samples for the experiment, cultured cells and mouse adipose tissue were collected and treated with lysis buffer (PRO-PREP; Intron Biotechnology, Seoul, Republic of Korea) for 60 min at 4Â°C. The extracted protein samples were between 35 and 40 Î¼g and subjected to SDS-PAGE on gels ranging from 7 to 12%, followed by transfer to nitrocellulose membranes (Amersham Biosciences, Westborough, MA, USA). Then, primary antibodies specific to the target protein were used to probe the membranes, followed by incubation with secondary antibodies conjugated to horseradish peroxidase (Santa Cruz Biotechnology, USA). Enhanced chemiluminescence kits (Amersham Biosciences) were used to detect signals.
The following antibodies were used for the experiments: anti-SCD1 (1:1,500), anti-SREBP1 (1:1,500), anti-phospho-PKA (1:1,000), anti-PKA (1:2,000), anti-phospho-p38 (1:1,000), anti-p38 (1:2,000), and anti-Î²-actin (1:2,500) from Santa Cruz Biotechnology and anti-phospho hormonesensitive lipase (HSL) (1:1,000) and anti-HSL (1:2,000) from Cell Signaling Technology (Danvers, MA, USA).

Sun J.L., Kim Y.J., Cho W., Park S.S., Abd El-Aty A.M., Mobarak E.H., Jung T.W, & Jeong J.H. (2024). The Extract of Humulus japonicus Inhibits Lipogenesis and Promotes Lipolysis via PKA/p38 Signaling. Obesity facts.

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Protein Expression Analysis by Western Blot

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Western blot was performed as previously described. Tissue and cell extracts were separated by electrophoresis on a 10% SDS-PAGE gel at 65 V for 45 min and 115 V for 1 h 20 min, then transferred to a PVDF membrane at 200 mA for 1 h. Membranes were blocked with 5% milk in TBST for 2 h and incubated overnight at 4 °C with the following primary antibodies: anti-MyD88, anti-α-SMA (Abcam, Cambridge, Cambs, UK), anti-FASN (Santa Cruz Biotechnology, TX, USA), anti-SREBP1 (Santa Cruz Biotechnology, TX, USA), anti-SCD1 (Santa Cruz Biotechnology, TX, USA), anti-STAT6 and anti-p-STAT6 antibodies (Affinity Biosciences, Liyang, Jiangsu, CN). HRP-conjugated goat anti-mouse IgG and goat anti-rabbit IgG were used as secondary antibodies. Blots were scanned using a Clinx Science Instrument. All specific bands were quantified with the ImageJ (version 1.8 RRID:SCR_003070) automated Digitizing System.

Liu Y., Chen H., Yan X., Zhang J., Deng Z., Huang M., Gu J, & Zhang J. (2024). MyD88 in myofibroblasts enhances nonalcoholic fatty liver disease-related hepatocarcinogenesis via promoting macrophage M2 polarization. Cell Communication and Signaling : CCS, 22, 86.

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