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Mab2510

Manufactured by Merck Group
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MAB2510 is a laboratory equipment product manufactured by Merck Group. It is designed for scientific research and analysis, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach. The capabilities and intended use of this product may vary, and further information would be required to present a concise and accurate description.

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Lab products found in correlation

Axio observer z1, by Zeiss (6 mentions) Lsm 710, by Zeiss (4 mentions) Ab66763, by Abcam (2 mentions) Prolong diamond antifade mountant, by Thermo Fisher Scientific (2 mentions) Ab47244, by Thermo Fisher Scientific (2 mentions) Ab11267, by Merck Group (2 mentions) Hoechst, by Merck Group (2 mentions) Mab2166, by Abcam (2 mentions) Pa5 34974, by Abcam (2 mentions) Ab15452, by Merck Group (2 mentions) Gfp trap gta 20, by Proteintech (1 mentions) P0044, by Merck Group (1 mentions) Hpa008352, by Atlas Antibodies (1 mentions) Nbt bcip, by AppliChem (1 mentions)

4 protocols using mab2510

1

Immunostaining of Neuronal Cells

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STHdh cells and primary cortical neurons were grown on poly-D-Lysine-coated coverslips. Cells were fixed with 4% paraformaldehyde for 15–20 min at room temperature. Cells were permeabilized with 0.1% Triton X-100 (in phosphate buffered saline, PBS), blocked with PBS containing 10% FBS and 0.05% Triton X-100 for 1h at room temperature. Primary antibodies were incubated overnight at 4°C in blocking solution with the following dilutions: HTT (Merck-Millipore #MAB2166, 1:100), PRMT2 (Abcam #ab66763, 1:100), PRMT6 (Abcam #ab47244, 1:100), mCherry (Life Technologies #PA5-34974, 1:600), MAP2 (Abcam #ab11267, 1:500 or Merck-Millipore #AB15452, 1:100), GFP (Merck-Millipore #MAB2510, 1:1000). The next day, after three washes with PBS, Alexa Fluor conjugated secondary antibodies (1:1000) were incubated for 1h at room temperature in the dark. The coverslips were then washed three times with PBS, incubated with Hoechst (Sigma #B2261) diluted in PBS for 10 min and mounted on glass slides with ProLong Diamond Antifade Mountant (Life technologies #P36961). Primary neurons and STHdh slides were imaged with a 63x oil-immersion objective using the Zeiss Axio Observer Z1 inverted microscope or an inverted confocal microscope (LSM 710, Zeiss) coupled to an Airyscan detector, respectively.
Migazzi A., Scaramuzzino C., Anderson E.N., Tripathy D., Hernández I.H., Grant R.A., Roccuzzo M., Tosatto L., Virlogeux A., Zuccato C., Caricasole A., Ratovitski T., Ross C.A., Pandey U.B., Lucas J.J., Saudou F., Pennuto M, & Basso M. (2021). Huntingtin-mediated axonal transport requires arginine methylation by PRMT6. Cell reports, 35(2), 108980.
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2

Immunostaining of Neuronal Cells

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STHdh cells and primary cortical neurons were grown on poly-D-Lysine-coated coverslips. Cells were fixed with 4% paraformaldehyde for 15–20 min at room temperature. Cells were permeabilized with 0.1% Triton X-100 (in phosphate buffered saline, PBS), blocked with PBS containing 10% FBS and 0.05% Triton X-100 for 1h at room temperature. Primary antibodies were incubated overnight at 4°C in blocking solution with the following dilutions: HTT (Merck-Millipore #MAB2166, 1:100), PRMT2 (Abcam #ab66763, 1:100), PRMT6 (Abcam #ab47244, 1:100), mCherry (Life Technologies #PA5-34974, 1:600), MAP2 (Abcam #ab11267, 1:500 or Merck-Millipore #AB15452, 1:100), GFP (Merck-Millipore #MAB2510, 1:1000). The next day, after three washes with PBS, Alexa Fluor conjugated secondary antibodies (1:1000) were incubated for 1h at room temperature in the dark. The coverslips were then washed three times with PBS, incubated with Hoechst (Sigma #B2261) diluted in PBS for 10 min and mounted on glass slides with ProLong Diamond Antifade Mountant (Life technologies #P36961). Primary neurons and STHdh slides were imaged with a 63x oil-immersion objective using the Zeiss Axio Observer Z1 inverted microscope or an inverted confocal microscope (LSM 710, Zeiss) coupled to an Airyscan detector, respectively.
Migazzi A., Scaramuzzino C., Anderson E.N., Tripathy D., Hernández I.H., Grant R.A., Roccuzzo M., Tosatto L., Virlogeux A., Zuccato C., Caricasole A., Ratovitski T., Ross C.A., Pandey U.B., Lucas J.J., Saudou F., Pennuto M, & Basso M. (2021). Huntingtin-mediated axonal transport requires arginine methylation by PRMT6. Cell reports, 35(2), 108980.
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3

Co-Immunoprecipitation of PAK4 and N-WASP

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H1299 cell were transfected with an EGFP vector or an EGFP-PAK4 vector by Lipofectamine 3000. Cells were harvested 48h after transfection and lysed with NP-40 cell lysis buffer containing 50 mM Tris-HCl pH7.5, 150 mM NaCl, 5 mM MgCl2, 1 mM EDTA, 10% Glycerol, 1% NP-40, and freshly added protease inhibitors (1697498, Roche) and phosphatase inhibitors (P0044, Sigma). 1000 μg protein lysate were immunoprecipated by GFP-Trap (gta-20, ChromoTek), CCTε and N-WASP antibodies with rabbit IgG as control, separated and probed with rat CCTε antibody (MCA2178, BIO-RAD), rabbit ARPC2 (HPA008352, Atlas Antibodies) and rabbit N-WASP antibody (HPA005750, Atlas Antibodies) and mouse GFP antibody (MAB2510, Milipore). For endogenous protein-protein interactions, 1000 μg protein lysate from MCF7 or H1299 cells were immunoprecipated by a rabbit anti-PAK4 antibody (6508) or by a rabbit anti-N-WASP antibody (HPA005750, Atlas Antibodies) using rabbit IgG as control. The immunoprecipitations were, separated on SDS-PAGE, blotted and probed with rabbit anti-N-WASP or anti-PAK4 antibodies.
Zhao M., Spiess M., Johansson H.J., Olofsson H., Hu J., Lehtiö J, & Strömblad S. (2017). Identification of the PAK4 interactome reveals PAK4 phosphorylation of N-WASP and promotion of Arp2/3-dependent actin polymerization. Oncotarget, 8(44), 77061-77074.
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4

Expression and Detection of Mutant ATXN1

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MSCs (5 × 105) were lysed in 1% SDS PBS containing protease inhibitors (Calbiochem) and benzonase (Novagen). Cell extracts were analyzed in an 8% SDS-PAGE. Ectopic expression of YFP-ATXN1(Q30) or (Q82) was validated by Western blotting with the polyclonal anti-ATXN1 SA4645 antibody [22 ] or anti-GFP antibody (MAB2510, Millipore). β-actin was detected with mouse monoclonal anti-β-actin antibody (sc-47778, Santa Cruz). After incubation with an appropriate alkaline-phosphatase conjugated secondary antibody, proteins were detected with NBT-BCIP (Applichem).
Laidou S., Alanis-Lobato G., Pribyl J., Raskó T., Tichy B., Mikulasek K., Tsagiopoulou M., Oppelt J., Kastrinaki G., Lefaki M., Singh M., Zink A., Chondrogianni N., Psomopoulos F., Prigione A., Ivics Z., Pospisilova S., Skladal P., Izsvák Z., Andrade-Navarro M.A, & Petrakis S. (2020). Nuclear inclusions of pathogenic ataxin-1 induce oxidative stress and perturb the protein synthesis machinery. Redox Biology, 32, 101458.
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