Newly Validated Antibodies for IHC and ICC

HeLa cells untreated (left panels) or treated with Nocodazole to arrest cell cycle at G2/M (right panels) were fixed and permeabilized with PFA and ice-cold methanol, and then intracellularly stained with Purified anti-Rb Phospho (Ser795) (clone A24001E)(bottom panels) or Purified Mouse IgG1, κ isotype control (Cat. No. 400101) (top panels) followed by Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322). Cells were counterstained with DAPI (Cat. No. 422801) (blue). Images were captured using a Revvity Operetta CLS™ High Content Analysis System with a 63X objective. Scale bar: 50 μm

A-431 cells untreated (top panels) or treated with ultraviolet light to induce DNA damage and p53 activation (bottom panels) were fixed and permeabilized with PFA and ice-cold methanol, and then intracellularly stained with either Alexa Fluor® 647 anti-p53 Phospho (Ser15) Recombinant (clone P53S15-1C11.Rec) (right panels) or Alexa Fluor® 647 Rabbit IgG isotype control (left panels). Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). Images were captured using a Revvity Operetta CLS™ High Content Analysis System with a 63X objective. Scale bar: 50 µm

IHC staining of Purified anti-Histone H3 (C-terminus) (clone 1B1-B2.recM1) on formalin-fixed paraffin-embedded human colon tissue. Following antigen retrieval using 1X Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422704), the tissue was incubated without (panel A) and with (panel B) Purified anti-Histone H3 (C-terminus) Recombinant (clone 1B1-B2.recM1) followed by incubation with Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322) for 1 hour at room temperature. The tissue was subsequently co-stained with Alexa Fluor® 488 anti-Vimentin (clone O91D3) (green) and nuclei were counterstained with DAPI (Cat. No. 422801) (blue). Images were captured with a 40X objective and merged. Scale bar: 50 μm

HeLa cells were fixed with 4% PFA Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 0.5% Triton X-100 for 10 minutes and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with Alexa Fluor® 488 anti-Giantin (clone Poly23A01) (panel A). Cells were also co-stained with Alexa Fluor® 647 anti-Tubulin-α (clone 10D8) (panel B) and nuclei were counterstained with DAPI (Cat. No. 422801) (panel C). Scale bar: 50 µm