Cells were fixed with 2% paraformaldehyde for 20 min at room temperature, followed by blocking with 0.1% BSA, 0.3% Triton X-100, 5% normal goat serum, in 1× PBS. Incubation with the primary antibodies was performed in blocking buffer and done overnight at 4 °C. The working solutions were as follows: rabbit anti-RLBP1 1:200 (PA5–29759, Thermo Fisher, Waltham, USA), rabbit anti-MITF 1:200 (PA5–38294, Thermo Fisher, Waltham, USA), rabbit anti-ZO1 1:100 (61–7300, Thermo Fisher, Waltham, USA), rabbit anti-BEST1 1:100 (ab14928, Abcam, Cambridge, UK). The immunoreactivity of the antibodies was confirmed by immunostainings on human retinal cryosections and ARPE19 cells as positive control (Fig. S1 ). As a secondary antibody we used the Alexa Fluor 594 goat-anti-rabbit 1:1000 (A-111012, Thermo Fisher, Waltham, USA). Cell nuclei were counterstained with DAPI (Thermo Fisher, Waltham, USA). Cells were imaged using a Leica TCS SP8 X confocal microscope.
Rabbit anti mitf
Manufactured by Thermo Fisher Scientific
Sourced in United States
Rabbit anti-MITF is a primary antibody that specifically targets the MITF (Microphthalmia-associated Transcription Factor) protein. MITF is a key transcription factor involved in the regulation of melanocyte development and function. The rabbit anti-MITF antibody can be used for the detection and analysis of MITF expression in various research applications.
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Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions)
Rabbit anti zo 1, by Thermo Fisher Scientific (2 mentions)
Tcs sp8 x confocal microscope, by Leica (1 mentions)
Alexa fluor 594 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Rabbit anti laminin, by Merck Group (1 mentions)
Confocal microscopy, by Leica (1 mentions)
Mouse anti sox10, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti k14, by Fortrea (1 mentions)
2 protocols using rabbit anti mitf
1
Immunocytochemistry of Retinal Cells
2
Immunostaining of Skin and Cell Cultures
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Skin cryosections or cultured cells were fixed in 4% paraformaldehyde for 1 h and permeabilized in 0.25% Triton X‐100 for 15 min at room temperature, blocked in 1% BSA for 30 min at room temperature and then stained with Mouse anti‐MyHC (Millipore, 05–716), Rabbit anti‐K14 (Covance, 905301), Mouse anti‐Sox10 (Santa Cruz Biotechnology, SC‐365692), Mouse anti‐Runx3 (R&D Systems, MAB3765‐SP), Rabbit anti‐Mitf (Thermo Fisher, MA5‐32554), Mouse anti‐Pax7 (DSHB), and Rabbit anti‐Laminin (Sigma, L9393) at 4°C overnight with gentle rocking, followed by Alexa 488‐, 561‐ or 647‐labelled anti‐mouse or anti‐rabbit secondary antibody (Invitrogen) staining at room temperature for 1 h. Nuclei were stained with 5 mg/mL DAPI (Invitrogen) for 5 min. All images were acquired by confocal microscopy (Leica). For Pax7 staining, heat‐activated antigen retrieval was performed by placing the paraformaldehyde‐fixed samples in citrate buffer (pH 6.0) at 95°C for 20 min and cooling the slides at room temperature for 20 min, followed by antibody incubation as described above.
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