Merlin

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Merlin is a versatile lab equipment designed for performing a range of scientific experiments. It is a compact and user-friendly device that offers precise control and monitoring of various parameters. The core function of the Merlin is to provide a reliable and consistent platform for researchers to conduct their studies.

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Anti-Merlin (3 citations) Merlin (sc-332, (2 citations)

5 protocols using merlin

Analyzing GLI1 and GLI2 Expression in NF2-Depleted Cells

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Cell lysates were collected in NP-40 buffer. Cytosolic and nuclear fractions were collected with an NE-PER kit (Pierce, Rockford, IL). Antibodies used include GLI1 (#2643; Cell Signaling, Danvers, MA), Merlin (#sc-55574; Santa Cruz Biotechnology, Santa Cruz, CA), GLI2 (#PA1941; Boster Bio Pleasanton, CA), GAPDH (#2118; Cell Signaling). Blots were developed with SuperSignal substrate (Pierce). The cytosolic and nuclear fractions were confirmed with anti-β-tubulin (#2146; Cell Signaling) or anti- HDAC1 (#2062; Cell Signaling) respectively. To assess the expression and localization of GLI1 and GLI2 in the MCF10AT KD cells (knockdown for NF2) in comparison to the MCF10AT NT (transfected with non-targeting vector), 100 μg of cytosolic and nuclear lysates were immunoprecipitated with either GLI1 or GLI2 antibodies and immunoblotted with the corresponding antibodies.

Das S., Jackson W.P., Prasain J.K., Hanna A., Bailey S.K., Tucker J.A., Bae S., Wilson L.S., Samant R.S., Barnes S, & Shevde L.A. (2017). Loss of Merlin induces metabolomic adaptation that engages dependence on Hedgehog signaling. Scientific Reports, 7, 40773.

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Immunofluorescence Analysis of GBM Markers

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Human GBM and normal brain tissues were obtained from the Cooperative Human Tissue Network (CHTN). Anti-FRMD6 (Sigma), -Lats1/2 (Bethyl Lab), -MST1/2, -YAP, -c-Met, -PDGFRA, -PDGFRB, and -merlin (Santa Cruz), -actin (Sigma), -v5 epitope (Invitrogen), -phospho-Lats1, -phospho-YAP (Cell signaling), -phospho-c-Met (Invitrogen and Santa Cruz), and -phospho-PDGFRA/B (Santa Cruz and R & D Systems) antibodies were used in the experiments. Anti-Ki67 was from the Fisher Scientific. Secondary anti-rabbit Alexa Fluor^® 594 and anti-mouse Alexa Fluor^® 488 were from Invitrogen.

Xu Y., Wang K, & Yu Q. (2016). FRMD6 inhibits human glioblastoma growth and progression by negatively regulating activity of receptor tyrosine kinases. Oncotarget, 7(43), 70080-70091.

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Western Blotting of Protein Extracts

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Western blots of protein extracts prepared from VS or GAN tissue or culture lysates were performed as described previously (Brown and Hansen 2008 (link); Hansen et al. 2006 (link); Yue et al. 2011 (link)). The primary antibodies used were anti-p75^NTR (kindly provided by Dr. Moses Chao), phosphorylated JNK (pJNK, Cell Signaling), JNK (Cell Signaling), phosphorylated JUN (pJUN, Cell Signaling), merlin (Santa Cruz), cleaved caspase-3 (Cell Signaling), RIP2 (Enzo Lifesciences, Farmingdale, NY), sortilin (Abcam), β-actin (Sigma), and Rho-GDI (Cell Signaling). Secondary antibodies (dilution,1:5000-50,000; Santa Cruz) were conjugated with horseradish peroxidase. Blots were developed using Super Signal West Femto kit (Thermo Fisher Scientific, Rockford, IL) and exposed to film (Amersham Hyperfilm TM ECL; GE Healthcare). As needed, membranes were stripped and re-probed with other antibody combinations. Densitometry to quantify protein levels was performed as previously described and statistical significance was determined with a student's non paired, two-tailed t- test.

Ahmad I., Yue W.Y., Fernando A., Clark J.J., Woodson E.A, & Hansen M.R. (2014). p75NTR is highly expressed in vestibular schwannomas and promotes cell survival by activating NF-κB. Glia, 62(10), 1699-1712.

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Immunoblotting of Sciatic Nerve Proteins

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Immunoblotting was performed as described in [32 (link)]. Separate pooled protein lysates were prepared from both intact and crushed sciatic nerves. The following primary antibodies were used: phospho-c-Jun (Ser73, 1:1000, Cell Signaling, USA, 8752), Neuregulin 1 (1:250, Santa Cruz, USA, clone C-20), Erk (1:500, Cell Signaling, USA), phospho-Erk (T202/Y204, 1:500, Cell Signaling, USA), ErbB2 (1:500, Cell Signaling, USA), GAPDH (1:1000, Santa Cruz, USA, 6C5) and merlin (1:500, Santa Cruz, USA A19). Results were quantified using gel analysis software by ImageJ. Density values were normalized to GAPDH and appropriate controls of transfection or wild-type tissue, respectively. In the case of phospho-specific detection of proteins, their acquired densities were referred to signals derived from related pan-antibodies that served as loading control (e.g., phospho-Erk to Erk signals).

Schulz A., Büttner R., Hagel C., Baader S.L., Kluwe L., Salamon J., Mautner V.F., Mindos T., Parkinson D.B., Gehlhausen J.R., Clapp D.W, & Morrison H. (2016). The importance of nerve microenvironment for schwannoma development. Acta Neuropathologica, 132, 289-307.

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Immunoblotting and Immunoprecipitation Analysis

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Immunoblotting analysis was performed as described previously^{25 (link)} using antibodies specific for β-catenin [Cell Signaling Technology (CST); 8480], CBP (CST; 7389), Merlin (Santa Cruz; sc-331), FOXM1 (Santa Cruz; sc-376471), p21 (CST; 2947), survivin (CST; 2808), and β-actin (CST; 4970). For immunoprecipitations, cell lysates and indicated antibodies (1 µg) were subjected to overnight rotation at 4°C. Protein A-Sepharose beads (40 µL, GE Healthcare) were added for 3 hours, followed by centrifugation, washed four times, and boiled with loading buffer for immunoblotting analysis.

Deng J., Hua L., Han T., Tian M., Wang D., Tang H., Sun S., Chen H., Cheng H., Zhang T., Xie Q., Wan L., Zhu H, & Gong Y. (2020). The CREB-binding protein inhibitor ICG-001: a promising therapeutic strategy in sporadic meningioma with NF2 mutations. Neuro-oncology Advances, 2(1), vdz055.

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