Anti rfp primary antibody

Manufactured by Rockland Immunochemicals

The Anti-RFP primary antibody is a laboratory reagent designed to detect and bind to the Red Fluorescent Protein (RFP) in biological samples. It is a purified polyclonal antibody produced by immunizing animals with RFP. The antibody can be used in various immunoassays and imaging techniques to identify and localize RFP-tagged proteins or cells.

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Lab products found in correlation

Axio imager, by Zeiss (2 mentions) Cm3050, by Leica (1 mentions) Donkey anti rabbit alexa 568, by Thermo Fisher Scientific (1 mentions) Slowfade gold, by Thermo Fisher Scientific (1 mentions) Lsm 700 confocal system, by Zeiss (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 secondary antibody, by Thermo Fisher Scientific (1 mentions) Plan apochromat 40 1.4na oil objective, by Zeiss (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Rnascope multiplex fluorescent reagent kit v2, by Advanced Cell Diagnostics (1 mentions)

2 protocols using anti rfp primary antibody

Transcardial Perfusion and Immunohistochemistry

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For general histology mice were anaesthetized with Euthatal (0.15–0.2 ml) and transcardially perfused with 10 ml of ice-cold PBS with heparin (0.02 mg ml−1) followed by 4% PFA in PBS solution. Brains were post-fixed overnight at 4 °C then transferred to PBS solution. Sections (50 μm) were cut with a cryostat (Leica CM3050S) and stained. Unless otherwise stated, sections were imaged with an epifluorescence microscope (Zeiss Axio Imager using ZEN 3.5 Blue Edition software). Immunohistochemistry was performed to enhance the signal of hM4D-mCherry-expressing RSP neurons. Sections were initially blocked using a 5% normal donkey serum (NDS) in PBS solution. Subsequently, they were incubated overnight at 4°C with anti-RFP primary antibody (1:1000, rabbit; 600-401-379, Rockland), followed by a 2-hour incubation with Alexa-568 donkey anti-rabbit (1:1000, Life Technologies). Antibodies were diluted in 0.5% NDS and 0.05% Triton X-100. DAPI was used for counterstaining. Sections were mounted in SlowFade Gold (Thermo Fisher, S36936) onto slides before imaging.

Campagner D., Vale R., Tan Y.L., Iordanidou P., Arocas O.P., Claudi F., Stempel A.V., Keshavarzi S., Petersen R.S., Margrie T.W, & Branco T. (2022). A cortico-collicular circuit for orienting to shelter during escape. Nature, 613(7942), 111-119.

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RNA Expression Profiling in Muscle Stem Cells

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In situ hybridization was performed using the RNAscope Multiplex Fluorescent Reagent kit v2 (Advanced Cell Diagnostics) following manufacturer's protocol with some modifications. TA from Pax7CRE-ER;ROSA26tdTomato mice was dissected and fixed in 2% PFA for 2 h at 4 ºC, frozen and processed into 10 μm longitudinal-sections. Slices were digested with Protease Plus. Probe hybridization was performed by incubation of mRNA target probes for 2 h at 40 °C. Slices were hybridized with two probe-sets of dual probes using Cd34-C3 as common probe in each set and Myog-C2 or Notch3-C2 as companion probes. Cd34-C3 probe was conjugated to Opal 690 fluorophore while Myog-C2 and Notch3-C2 were conjugated to Opal 570 fluorophore (Akoya Biosystems). For SC immunofluorescence, sections were blocked in 3% BSA for 1 h at room-temperature prior to incubation overnight with anti-RFP primary antibody (Rockland, 600-401-379, 1:200) at 4 ºC. Anti-rabbit Alexa Fluor 488 secondary antibody (A11008, Life Technologies, 1:500) and DAPI (Invitrogen) were added for 1 h at room-temperature. After washing, sections were mounted with Prolong Gold Antifade Mountant (Thermo Fisher). Confocal images were taken with Zeiss LSM-700 confocalsystem with a Plan-Apochromat 40 × /1.4 NA oil objective.

García-Prat L., Perdiguero E., Alonso-Martín S., Dell'Orso S., Ravichandran S., Brooks S.R., Juan A.H., Campanario S., Jiang K., Hong X., Ortet L., Ruiz-Bonilla V., Flández M., Moiseeva V., Rebollo E., Jardí M., Sun H.W., Musarò A., Sandri M., Del Sol A., Sartorelli V, & Muñoz-Cánoves P. (2020). FoxO maintains a genuine muscle stem-cell quiescent state until geriatric age. Nature cell biology, 22(11).

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