Dulbecco’s modified minimal essential medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS), Trypsin-EDTA solution, 100× penicillin–streptomycin–l -glutamine solution, anti-RFP antibody, Alexa 647-conjugated secondary antibody, SlowFade Gold antifade reagent, and a LIVE/DEAD fixable yellow dead cell stain kit were purchased from Thermo Fisher (Waltham, MA USA). Puromycin solution and G418 solution were obtained from InvivoGen (San Diego, CA, USA). Hygromycin B solution was purchased from Enzo Life Science (Farmingdale, NY, USA). Anti-ORF57, anti-K8, and anti-K8.1 antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Anti-LANA and anti-lamin A/C antibodies were purchased from Millipore-Sigma (St. Louis, MO, USA). Anti-nucleolin antibody was purchased from Abcam (Cambridge, UK). Anti-K-Rta antibody was described previously (68 (link)). Monoclonal anti-ORF52 antibody was a generous gift from Fanxiu Zhu (Florida State University). All other chemicals were purchased from Millipore-Sigma (St. Louis, MO, USA) unless otherwise stated.
Anti rfp antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-RFP antibody is a laboratory reagent used to detect and quantify the expression of Red Fluorescent Protein (RFP) in various experimental systems. It is a highly specific and sensitive tool for researchers studying gene expression, protein localization, and other applications involving RFP-tagged proteins.
Automatically generated - may contain errors
Lab products found in correlation
Alexa 647 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Slowfade gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Trypsin edta solution, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Live dead fixable yellow dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified minimal essential medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin l glutamine solution, by Thermo Fisher Scientific (1 mentions)
Hygromycin b solution, by Enzo Life Sciences (1 mentions)
Anti orf57, by Santa Cruz Biotechnology (1 mentions)
Anti k8, by Santa Cruz Biotechnology (1 mentions)
Puromycin solution, by InvivoGen (1 mentions)
Anti nucleolin antibody, by Abcam (1 mentions)
Alexa conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Anti phosphotyrosine 4g10, by Merck Group (1 mentions)
Anti socs3 l210, by Cell Signaling Technology (1 mentions)
0.2 μm nitrocellulose membrane, by GE Healthcare (1 mentions)
Edta free protease inhibitor cocktail, by Roche (1 mentions)
β actin antibody, by Fortis Life Sciences (1 mentions)
Chemidoc xrs software image lab 5, by Bio-Rad (1 mentions)
4 protocols using anti rfp antibody
1
Immunofluorescence Staining Protocol
2
Antibody Acquisition for Protein Analysis
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Anti-FLAG (SIG1–25), anti-HA (HA-7), and anti-β-tubulin (SDL.3D10) antibodies were purchased from Sigma-Aldrich. The anti-RFP antibody was from Thermo Fisher Scientific. Anti-phosphotyrosine (4G10) and anti-NeuN (A60) antibodies were from Merck Millipore. The anti-PTPRJ (AF1934) antibody was from R&D Systems. Anti-SH2B1 (E-20) and anti-OB-R (B-3) antibodies were from Santa Cruz Biotechnology. Anti-JAK2 (D2E12), anti-STAT3 (124H6), anti-phosphorylated STAT3 (phospho Y805, D3A7), and anti-SOCS3 (L210) antibodies were from Cell Signaling Technologies. The anti-phosphorylated JAK2 (phospho Y1007 + Y1008, E132) antibody was from Abcam. Anti-NPY (22940) and anti-α-MSH (20074) antibodies were from Immunostar. HRP-conjugated and Alexa-conjugated secondary antibodies were from Thermo Fisher Scientific.
3
Western Blot Analysis of Signaling Proteins
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A549 cells were lysed using 1%-Triton X-100 (pH 7.4), EDTA-free protease inhibitor cocktail (Roche Diagnostics) in PBS. Cell lysates were subjected to SDS-PAGE and separated proteins were transferred onto 0.2 μm nitrocellulose membrane (GE Health care) and blotted with specific antibodies. β-catenin and FLAG antibodies were purchased from Sigma-Aldrich. LRP5 antibody was obtained from Cell signaling. β-actin antibody was purchased from Bethyl Laboratories. HBD3-8A antibody was deposited to the DSHB by Starner, T. (DSHB Hybridoma Product hBD-3-8A). An anti-RFP antibody was purchased from Invitrogen. Western blots were quantified using ChemiDoc™ XRS + software Image Lab 5.1 (BioRad).
4
Immunostaining of Mouse Retina and Organoids
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For analyzing cross-sectional images of mouse NR and RPE, the eye was enucleated, the cornea was punctured, and the eye was fixed for 1 h in 4% paraformaldehyde in PBS. The fixed eyes were cryopreserved by sinking in 15% sucrose in PBS, followed by equilibration in 30% sucrose. For analyzing cross-sectional images of retinal organoid, the organoid was fixed in 1% paraformaldehyde in PBS. The fixed organoids were cryopreserved by sinking in 10% sucrose in PBS, followed by equilibration in 30% sucrose. Mouse eyes and retinal organoids were embedded in OCT compound and snap-frozen in liquid nitrogen. Samples were cut into 7-μm thick sections using a cryostat. Cryosections of mouse retina were immunostained with the primary antibody, anti-RFP antibody (Invitrogen, MA, USA), and secondary antibody, Alexa Fluor™ 594 anti-rabbit IgG (Invitrogen, MA, USA). Retinal organoids were immunostained with various markers of retinal cell types according to the developmental stage. The antigen, host species, used concentration, and supplier of the primary antibody used are listed in Supplementary Table 2.
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