goat anti human igg  (R&D Systems)

Journal: The FASEB Journal

Article Title: Loss of the RNA Binding Protein HuR in Early Murine Limb Mesenchyme Does Not Affect Development but Leads to Impaired Bone Homeostasis in Adulthood

doi: 10.1096/fj.202500780RR

Figure Lengend Snippet: Effect of germline Cre recombination in HuR fl/fl mice (Elavl1 KO) through maternal inheritance of Prx1‐Cre. (A) Whole‐mount embryos at E13.5 stained with alcian blue. (B) Toluidine blue‐stained sections of E13.5 embryo forelimbs and hindlimbs. (C) Immunofluorescence detection of HuR in limb sections from E13.5 embryos. (D) Brightfield micrographs demonstrating morphology changes in Control and Elavl1 KO embryos isolated from E9.5 to E11.5. Scale bar = 500 μm. (E) Whole‐mount in situ hybridization chain reaction detection of limb bud markers mRNA in E10.5 and E11.5 embryos. Asterisk: Limb bud, Triangle: Apical ectodermal ridge, Arrowhead: The zone of polarizing activity. Scale bar = 150 μm. (F) Whole‐mount immunofluorescence detection of Fgf8 protein in E10.5 embryos. Asterisk: Limb bud, Triangle: Apical ectodermal ridge. Scale bar = 150 μm.

Article Snippet: Embryos were fixed in 4% formaldehyde for 1 h at 4°C, washed, and stored in 0.0025% Triton X‐100 in PBS (PBS‐Tr) at 4° C. Prior to immunofluorescence staining, embryos were permeabilized with 0.25% Triton X‐100 in PBS for 30 min and then blocked with 2% bovine serum albumin in PBS solution for 1 h before incubation with 1:100 Fgf8 antibody (MAB323, R&D Systems, UK) overnight at 4°C.

Techniques: Staining, Immunofluorescence, Control, Isolation, In Situ Hybridization, Activity Assay