Sc 20158

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-20158 is a laboratory equipment product from Santa Cruz Biotechnology. It is designed for specific research applications. The core function of this product is to enable researchers to conduct their investigations effectively. However, a detailed and unbiased description of the product's features and specifications is not available.

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3 protocols using sc 20158

Protein Extraction and Western Blot Analysis

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Protein extracts from the heart and liver tissue were obtained by homogenization in lysis buffer (100 mg/mL) as mentioned previously [1 (link), 26 (link)]. The protein concentrations were determined by Lowry protein assay and the samples were electrophoresed in a 12% SDS polyacrylamide gel electrophoresis (SDS-PAGE), the proteins were then transferred to PVDF membranes (GE Healthcare Life Sciences, Pittsburgh, PA, USA). The membranes were blocked using 5% non-fat milk for 1 h. Monoclonal primary antibodies were diluted 1:1000 in antibody binding buffer (TBS) and used for hybridization overnight (4 °C) with the following antibodies ANP, phosphor-Akt, β-Actin, Bax, BNP, Cytochrome C, phosphor-GATA4, PGC1α (sc-20158, sc-7985, sc-47778, sc-526, sc-18818, sc-13560, sc-32823-R, sc-13067, Santa Cruz); phosphor-AMPK, Cle-Caspase-3, phosphor-FOXO3a, Sirt1 (#2535, #9664, #9466, #9475s, Cell Signaling, The Netherlands). Following hybridization with appropriate secondary antibodies the membranes were washed in for 10 min thrice. The blots were detected in chemiluminescent detection using ECL with Fujifilm LAS-3000 (GE Healthcare Life Sciences).

Asokan S.M., Wang T., Wang M.F, & Lin W.T. (2020). A novel dipeptide from potato protein hydrolysate augments the effects of exercise training against high-fat diet-induced damages in senescence-accelerated mouse-prone 8 by boosting pAMPK / SIRT1/ PGC-1α/ pFOXO3 pathway. Aging (Albany NY), 12(8), 7334-7349.

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Quantification of PVN Protein Levels

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The tissue homogenate from the PVN was subjected to a western blot analysis for determination of atrial natriuretic peptide (ANP) (sc-20158, Santa Cruz Biotechnology, Santa Cruz, CA, USA), β-myosin heavy chain (β-MHC) (sc-15929, Santa Cruz Biotechnology, Santa Cruz, CA, USA), TH (sc-14007, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and GAD67 (sc-7512, Santa Cruz Biotechnology, CA, USA) protein levels as previously described.^{32 (link)} We use molecular weight markers (Thermo Scientific, USA) to determine the location of each protein. Protein loading was controlled by normalizing all protein intensities to that of β-actin using a β-actin antibody (Thermo Scientific, USA). Band densities were analyzed using NIH ImageJ software.

Liang B., Zhao Y.N., Wang X., Yu X.J., Li Y., Yang H.Y., Su Q., Kang Y.M, & Yang Z.M. (2018). Angiotensin-(1-7) attenuates hypertension and cardiac hypertrophy via modulation of nitric oxide and neurotransmitter levels in the paraventricular nucleus in salt-sensitive hypertensive rats. RSC Advances, 8(16), 8779-8786.

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Comprehensive Antibody Detection Protocol

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Antibodies against CCR9 (ab38567, 1:500 dilution) were obtained from Abcam. Antibodies against the following proteins were purchased from Cell Signaling Technology: MEK1/2 (9122, 1:1000 dilution), phosphorylated (p) MEK1/2Ser32/36 (9154, 1:1000 dilution), ERK1/2 (4695, 1:1000 dilution), p‐ERK1/2Thr202/Tyr204 (4370, 1:1000 dilution), JNK (9252, 1:1000 dilution), p‐JNKThr183/Tyr185 (4668, 1:1000 dilution), P38 (9212, 1:1000 dilution), p‐P38Thr180/Tyr182 (4511, 1:1000 dilution), AKT (4691, 1:1000 dilution), p‐AKTSer473 (4060, 1:1000 dilution), glycogen synthase kinase 3β (GSK3β; 9315, 1:1000 dilution), p‐GSK3βSer9 (9322, 1:1000 dilution), p70S6 kinase (2708, 1:1000 dilution), p‐p70S6 kinaseSer371 (9208, 1:1000 dilution), mTOR (2983, 1:1000 dilution), p‐mTORSer2448 (2971, 1:1000 dilution), PI3 kinase p85 (4257, 1:1000 dilution), and p‐PI3 kinase p85Tyr458 (4228, 1:1000 dilution). Antibodies against β‐myosin heavy chain (sc53090, 1:200 dilution) and atrial natriuretic peptide (sc20158, 1:200 dilution) were purchased from Santa Cruz Biotechnology. Antibodies against GAPDH (MB001, 1:10 000 dilution) were obtained from Bioworld Technology. The BCA protein assay kit was purchased from Pierce (Thermo Fisher Scientific). We used peroxidase‐conjugated secondary antibodies (1:10 000 dilution; Jackson ImmunoResearch Laboratories) for visualization.

Xu Z., Mei F., Liu H., Sun C, & Zheng Z. (2016). C‐C Motif Chemokine Receptor 9 Exacerbates Pressure Overload–Induced Cardiac Hypertrophy and Dysfunction. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 5(5), e003342.

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