Alexa fluor 488 labeled goat anti mouse igg antibodies

Mice were sacrificed and spleens were isolated. Formalin-fixed, paraffin-embedded samples were segmented and stained with hematoxylin and eosin (HE). The pathologic changes of spleen were observed by the optical microscope (Nikon). The tissue sections (4 μm) were deparaffinized and rehydrated through xylene and graded alcohols. Tissue sections were boiled in citrate buffer for antigen retrieval, and blocked with 1% serum goat for 60 min. The slides were stained with the primary antibody anti-CD45 (Santa Cruz) at 4 °C overnight and the secondary Alexa Fluor 488-labeled goat anti-mouse IgG antibodies (Beyotime, China) at room temperature for 2 h. Nuclei were counterstained with DAPI. Images were acquired using a Nikon A1R confocal laser scanning microscope.

In Fig. 2e and Supplementary Fig. 2c, six-day-old 35 S::Flag-RTFL18 seedlings grown in WL were fixed in FAA solution (Gefanbio, M072). Seedlings were washed by 1 × PBS. Add enzymatic solution (10 mg/ml cellulase R-10, 2.5 mg/ml Macerozyme R-10, 0.4 M Mannitol, 20 mM MES pH = 5.7, 20 mM KCl, 10 mM CaCl₂) and shake for 2 h. Seedlings were washed by 1 × PBS. Seedlings were treated for 1 h in a 5% BAS solution containing 50 mM glycine. Flag-RTFL18 was detected via anti-Flag (GNI, GNI4110-FG) antibody as a primary antibody and Alexa Fluor 488-labeled Goat Anti-Mouse IgG antibodies as a secondary antibody (Beyotime, A0428). Fluorescence was imaged under a confocal laser scanning microscope (OLYMPUS FV3000).

The following antibodies were purchased: anti-Myc antibody (GNI, GNI4410-MC, 1:4000); anti-Flag antibody (GNI, GNI4410-FG, 1:4000); anti-H + -ATPase antibody (PHYTOAB, PHY2285A, 1:2000); anti-Rbcl antibody (Agrisera, AS03037, 1:4000); anti-H3 antibody (Abmart, P30266M, 1:3000); anti-β-Tubulin antibody (Abmart, M30109M, 1:4000); anti-GFP antibody (Abmart, M20004, 1:3000); anti-PIF4 antibody (Abiocode, R2534-4, 1:2000); anti-GST antibody (Abmart, M20007, 1:4000); anti-His antibody (GNI, GNI4110-HS, 1:4000); Alexa Fluor 488-labeled Goat Anti-Mouse IgG antibodies (Beyotime, A0428, 1:200); Glutathione Resin (GenScript, L00206); anti-GFP Affinity beads (Smart-Lifesciences, SA070001); rProtein A Beads 4FF (Smart-Lifesciences, SA015005); Ni NTA Beads 6FF (Smart-Lifesciences, SA005025).