rabbit anti-gfp a11122  (Thermo Fisher)

Journal: bioRxiv

Article Title: Neuronal feedback loop in the suprachiasmatic nucleus generates robust circadian rhythms

doi: 10.1101/2025.05.21.655394

Figure Lengend Snippet: (A) Schematic diagram showing the injection of AAV- EF1α-DIO-GFP::TeNT or AAV- CAG-FLEX-EGFP into the SCN of Avp-Cre mice. (B) Representative coronal SCN sections of mice with GFP::TeNT (immunostained with anti-GFP antibody) or EGFP expression. Scale bar, 200 μm. (C) Representative locomotor activity actograms of Avp-TeNT or Avp-EGFP (control) mice. Animals were initially housed in LD and then in DD. Gray shading indicates the time when the light was off. (D and E) The free-running period and amplitude (Qp values) of the locomotor activity rhythm in DD. Values are mean ± SEM; n = 15 for Avp-TeNT , n = 9 for Avp-EGFP mice. ***p < 0.001 by two-tailed Welch t test; ns, not significant.

Article Snippet: Antibodies used were rabbit anti-GFP antibody (1:1,000; A11122, Thermo Fisher Scientific) and rabbit anti-VIP antibody (1:1,000; #20077, Immunostar).

Techniques: Injection, Expressing, Activity Assay, Control, Two Tailed Test

Journal: Nature Communications

Article Title: The immune NIK1/RPL10/LIMYB signaling module regulates photosynthesis and translation under biotic and abiotic stresses.

doi: 10.1038/s41467-025-59571-y

Figure Lengend Snippet: a The bacterial PAMP, flg22, and viral PAMP, InRNA, induce phosphorylation of endogenous LIMYB. Col-0 seedlings were incubated with RNA prepared from begomovirus-infected leaves (InRNA), mock-inoculated leaves (UnRNA), H 2 O, and 100 mM flg22 for 3 h. LIMYB was immunoprecipitated from total protein extracts with an anti-LIMYB polyclonal antibody and immunoblotted with α-phosphoserine (α-pSer) (top) and α-LIMYB (bottom) antibodies. b Viral PAMP-induced LIMYB phosphorylation requires the NIK1/NIK2 function. limyb and nik1/nik2 protoplasts transfected with LIMYB-GFP were treated with InRNA or UnRNA, and the immunoprecipitated LIMYB-GFP was probed with an α-phosphoserine (α-pSer) antibody. c Flg22-induced phosphorylation of LIMYB requires the NIK1/NIK2 function. Col-0 and nik1/nik2 protoplasts transfected with LIMYB-GFP were treated with flg22. Half of the immunoprecipitated LIMYB was fractionated by SDS-PAGE and immunoblotted with an α-phosphoserine (α-pSer) antibody, and the other half with an α-GFP antibody . d LIMYB-S157A does not mediate the repression of the RPL18 promoter in the presence of InRNA. Col-0 protoplasts, LIMYB-S157A-expressing limyb protoplasts, and nik1nik2 protoplasts were transfected with plasmids carrying the RPL13 promoter fused to luciferase and treated with UnRNA or InRNA. After 16 h, luciferase activity was measured from total protein extracts of transfected protoplasts. Data are mean values ± SD ( n = 3) (two-tailed unpaired Student’s t -test, * p < 0.01, ns = not significant). e Ser157 is a possible phosphorylation site on LIMYB. The limyb protoplasts transfected with LIMYB-GFP or LIMYB-S157A-GFP were treated with InRNA or UnRNA and protein phosphorylation was assayed by immunoblotting immunoprecipitated GFP fusions with anti-phosphoserine antibody. f UnRNA fails to cause downregulation of NIK1 signaling-associated marker genes in LIMYB-S157A-expressing limyb protoplasts. RNA was extracted from UnRNA and InRNA-treated protoplasts, and transcript accumulation was determined by RT-qPCR. Results are mean values ± SD ( n = 3) (two-tailed unpaired Student’s t -test, * p < 0.01, ns = not significant). All the above experiments were repeated at least twice with similar results.

Article Snippet: LIMYB-GFP targets from LIMYB-32-L1-, LIMYB-32-L3-, and LIMYB-32-L4-isolated nuclei were immunoprecipitated using rabbit polyclonal anti-GFP - IgG antibody (Thermo Fisher Scientific, MA, USA, cat. # A11122, 1:5,000) and captured with protein-A agarose beads (Invitrogen TM , Cat # 15918014).

Techniques: Phospho-proteomics, Incubation, Infection, Immunoprecipitation, Transfection, SDS Page, Expressing, Luciferase, Activity Assay, Two Tailed Test, Western Blot, Marker, Quantitative RT-PCR