ChromoTek pan-RFP Polyclonal antibody

Western blot detection of fluorescent proteins from different lineages. SDS-PAGE was performed with 20 ng of purified recombinant proteins per lane. Positive signal could be obtained with red fluorescent proteins from Discosoma dsRed lineage (mCherry, dsRed, tdTomato), with synthetic red fluorescent protein mScarlet, with fluorescent proteins from Entacmaea lineages (mRuby2, mKate2, TagRFP, TagBFP). Purified recombinant green fluorescent proteins EGFP, TurboGFP and mNeonGreen were used as negative controls. Pan-RFP (pabr1) antibody was applied at 1:1000 dilution o/n at +4°C. Secondary antibody: goat anti-rabbit HRP. Note: Red fluorescent proteins often demonstrate multiple bands on Western blots due to partial fragmentation. These bands correspond to differently truncated forms of red fluorescent proteins.

Immunostaining of HeLa cells, transiently transfected with different fluorescent proteins. Primary antibody: pan-RFP (pabr1) 1:800  for 1 h RT. Secondary: goat anti-rabbit. Upper row shows the innate signals from transfected fluorescent proteins, nuclei are in cyan. Lower row shows the signals from immunostainings with pan-RFP (pabr1) antibody, nuclei are in cyan. Pan-RFP (pabr1) can be used for IF detection of mScarlet, mCherry, tdTomato, mPlum, TagRFP, mKate TagBFP, mRuby2.

Western blot detection of purified mCherry red fluorescent protein down to 1-2 ng. A serial dilution of purified recombinant mCherry protein (250 ng - 1 ng) was subjected to SDS-PAGE. Purified recombinant EGFP (250 ng) was used as negative control (last lane). Pan-RFP (pabr1) antibody was applied at 1:1000 dilution o/n at +4°C. Secondary antibody: goat anti-rabbit HRP.