Mertk

Manufactured by Cell Signaling Technology

Sourced in France, United States

MERTK is a protein tyrosine kinase that plays a role in the phagocytosis of apoptotic cells. It is involved in the clearance of dying cells and the regulation of the inflammatory response.

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Anti-Mertk antibody (2 citations)

5 protocols using mertk

Immunoblotting and Flow Cytometry Antibody Panel

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Antibodies to Cbl-b (#9498), Granzyme B (#17215), perforin (#62550) and Mertk (#4319) for immunoblotting analysis were purchased from Cell Signaling Technology (CST) and p-Mertk (#ab14921) from Abcam. An antibody to β-actin (#MAB1501R) for immunoblotting analysis was purchased from Millipore-Sigma. For flow cytometric analysis, the anti-hCD56-APC antibody (#B46024) was purchased from Beckman Coulter, anti-hCD3 (#130-113-134) and anti-hIFN-γ (#130-113-493) from Miltenyi Biotec, and anti-hTyro3 (FAB859P), anti-hAxl (#FAB154P) and anti-hMertk (FAB8912P) from R&D. The scrambled and Cbl-b-specific Accell siRNAs were purchased from Dharmacon. Recombinant human IL-2 and IL-15 proteins were obtained from National Institutes of Health (NIH). Recombinant human IL-7, IL-12, IL-18 and IL-21 were purchased from Miltenyi Biotec.

Lu T., Chen L., Mansour A.G., Yu M.J., Brooks N., Teng K.Y., Li Z., Zhang J., Barr T., Yu J, & Caligiuri M.A. (2021). Cbl-b is upregulated and plays a negative role in activated human NK cells. Journal of immunology (Baltimore, Md. : 1950), 206(4), 677-685.

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Western Blot Analysis of Tumor Lysates

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Whole cell lysates from tumors (12 ) or cell lines were prepared as previously described (11 (link)) and detailed in Suppl. Methods. Proteins were detected with the following antibodies: H3K27me3, H3K36me3, Histone H3, MYCN, MERTK, phospho ERK (all from Cell Signaling Technology) or β-actin (Sigma-Aldrich, #A2228).

Toledo R.A., Qin Y., Cheng Z.M., Gao Q., Iwata S., Silva G.M., Prasad M.L., Ocal I.T., Rao S., Aronin N., Barontini M., Bruder J., Reddick R.L., Chen Y., Aguiar R.C, & Dahia P.L. (2015). Recurrent mutations of chromatin remodeling genes and kinase receptors in pheochromocytomas and paragangliomas. Clinical cancer research : an official journal of the American Association for Cancer Research, 22(9), 2301-2310.

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Immunoblotting Analysis of Cellular Proteins

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Cell lysates were prepared and immunoblotting was carried out as previously described.^{13 (link)} Anti-TYRO3 (Ref. 5585), AXL (Ref. 8661), MERTK (Ref. 4319), caspase-8 (Ref. 9746), cleaved caspase-3 (Ref. 9664), cleaved PARP (Ref. 5625), FOXM1 (Ref. 5436), pRB (Ref. 9309), phospho-pRB Ser780 (Ref. 9307), CYCLIN D1 (Ref. 2926), AURORA A (Ref. 14475), AURORA B (Ref. 3094), SURVIVIN (Ref. 2808), c-MYC (Ref. 9402), anti-mouse IgG, HRP-linked (Ref. 7076) and anti-rabbit IgG, HRP-linked (Ref. 7074) antibodies were purchased from Cell Signaling Technology (Ozyme, Montigny-le-Bretonneux, France). Anti-α-TUBULIN (Ref. T6199) and anti-β-ACTIN (Ref. A2228) antibodies were obtained from Sigma-Aldrich. For peptide-N-glycosidase F (PNGaseF), we digested 10 µg of protein with PNGaseF, according to the manufacturer’s instructions (New England Biolabs, Evry, France).

Dufour F., Silina L., Neyret-Kahn H., Moreno-Vega A., Krucker C., Karboul N., Dorland-Galliot M., Maillé P., Chapeaublanc E., Allory Y., Stransky N., Haegel H., Menguy T., Duong V., Radvanyi F, & Bernard-Pierrot I. (2019). TYRO3 as a molecular target for growth inhibition and apoptosis induction in bladder cancer. British Journal of Cancer, 120(5), 555-564.

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Immunofluorescence Assays of RPE Cell Markers

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The immunofluorescence assays were performed as previously described in Feng et al. (64 (link)). HsRPE cells were fixed with 4% paraformaldehyde for 15 min, followed by blocking with 5% BSA at room temperature for 1 h. The cell samples were incubated with RPE65 (Abcam, ab235950), F-actin (Abcam, ab130935), CCT5 (Proteintech, 11603-1-AP), and MERTK (Cell Signaling Technology, 4319S) antibodies. Cell samples were then incubated with secondary antibodies for 1 h at room temperature. Subsequently, the nuclei of the cells were stained using DAPI for 7 min. All the images were taken using a confocal microscope (Carle ZEISS LSM 980, Oberkohen, Germany).

Feng L., Li H., Du Y., Zhang T., Zhu Y., Li Z., Zhao L., Wang X., Wang G., Zhou L., Jiang Z., Liu Z., Ou Z., Wen Y, & Zhuo Y. (2022). Chaperonin-Containing TCP1 Subunit 5 Protects Against the Effect of Mer Receptor Tyrosine Kinase Knockdown in Retinal Pigment Epithelial Cells by Interacting With Filamentous Actin and Activating the LIM-Kinase 1/Cofilin Pathway. Frontiers in Medicine, 9, 861371.

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Western Blot Analysis of Receptor Tyrosine Kinases

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Proteins were separated on an 8% SDS/PAGE gel and transferred onto a PVDF membrane. After blocking with 5% bovine serum albumin (BSA; Bio‐Rad, Hercules, CA, USA) for 1 h at room temperature, membranes were incubated with primary antibodies at 4 °C overnight. Antibody against GAPDH was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against AXL, p‐AXL, TYRO3, MERTK, EGFR, p‐EGFR, MET, p‐MET, p‐AKT, p‐ERK, and ERK were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibody against AKT was purchased from GeneTex Inc. (Irvine, CA, USA). The membranes were then incubated with horseradish peroxidase‐conjugated secondary antibodies, and proteins were detected using ECL reagents (GE Healthcare Life Sciences, Piscataway, NJ, USA).

Liao Y., Chuang Y., Lin H., Lin N., Hsu T., Hsieh S., Chen S., Hung J., Yang H., Liang J., Huang M, & Huang J. (2022). GALNT2 promotes invasiveness of colorectal cancer cells partly through AXL. Molecular Oncology, 17(1), 119-133.

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