Anti GFP

Immunofluorescence Microscopy of Anti-GFP (GOAT) Antibody. Tissue: E5.5 Hex-GFP transgenic mouse embryo. Primary antibody: Goat anti-GFP was used at 1:500 dilution. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody at 1:10,000 for 45 min at RT. Staining: GFP as green fluorescent signal with DAPI blue counterstain.

Immunofluorescence Microscopy of GFP-GOAT Antibody. Tissue: Drosophila melanogaster late stage embryonic central nervous system. Fixation: 0.5% PFA. Antigen retrieval: not required.Primary antibody: Anti-GFP antibody at a 1:1,000 for 1 h at RT.Secondary antibody: AlexaFluor 488？ conjugated Anti-Goat antibody at 1:300 for 45 min at RT.Panel A: shows a lateral view (ventral left). Panels B and C: shows ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up.Staining: tau-GFP cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) as green fluorescent signal.

Western Blot of Goat anti-GFP antibody. Lane 1: HeLa cells. Lane 2: mock transfected HeLa cell lysate. Load: 35 μg per lane. Primary antibody: GFP antibody at 1 μg/ml for 1 h at room temperature. Secondary antibody: IRDyeR 800 conjugated Donkey-a-Goat IgG [H&L] MX7 (605-732-125) secondary antibody at 1:2,500 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 27 kDa, 33 kDa for GFP. Other band(s): none.

Immunofluorescence Microscopy of GFP-GOAT-Antibody. Tissue: Sf-1:Cre mice crossed to the Z/EG reporter line. Mouse brain (coronal view, 20X magnification). Fixation: 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Antigen retrieval: frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40 microns. Primary antibody: Goat anti-GFP was used at 1:500 dilution in free floating imunnohistochemistry to detect GFP. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody was used for detection at 1:500 at 1:10,000 for 45 min at RT. Localization: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus. Staining: eGFP as green fluorescent signal and sections were counterstained with DAPI.

Multi-lysate Western Blot of Goat anti-GFP antibody. Marker: Opal Pre-stained ladder (p/n MB-210-0500). Lane 1: HEK293 lysate (p/n W09-000-365). Lane 2: HeLa Lysate (p/n W09-000-364). Lane 3: CHO/K1 Lysate (p/n W07-000-357). Lane 4: MDA-MB-231 (p/n W09-001-GK6). Lane 5: A431 Lysate (p/n W09-000-361). Lane 6: Jurkat Lysate (p/n W09-001-370). Lane 7: NIH/3T3 Lysate (p/n W10-000-358). Lane 8: E-coli HCP Control (p/n 000-001-J08). Lane 9: FLAG Positive Control Lysate (p/n W00-001-383). Lane 10: Red Fluorescent Protein (p/n 000-001-379). Lane 11: Green Fluorescent Protein (p/n 000-001-215). Lane 12: Glutathione-S-Transferase Protein (p/n 000-001-200). Lane 13: Maltose Binding Protein (p/n 000-001-385). Load: 10 μg of lysate or 50ng of purified protein per lane. Primary antibody: GFP antibody at 1ug/mL overnight at 4C. Secondary antibody: Peroxidase goat secondary antibody at 1:30,000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS (p/n MB-082) for 30 min at RT. Predicted/Observed size: 30 kDa for GFP.

Western Blot of Goat anti-GFP antibody with lysates. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: GFP/HEK293T WCL (p/n 000-001-215/W09-001-GX5) [0.05/10μg]. Lane 3: HEK293T WCL (p/n W09-001-GX5) [10μg]. Lane 4: GFP/NIH-3T3 WCL (p/n 000-001-215/W10-000-358) [0.05/10μg]. Lane 5: NIH-3T3 WCL (p/n W10-000-358) [10μg]. Lane 6: GFP/PC-12 WCL (p/n 000-001-215/W12-001-GL9) [0.05/10μg]. Lane 7: PC-12 WCL (p/n W12-001-GL9) [10μg]. Primary antibody: Anti-GFP antibody at 1ug/mL overnight at 2-8°C. Secondary antibody: Donkey Anti-Goat IgG HRP secondary antibody (p/n 605-703-002) at 1:40,000 for 30 min at RT. Blocking Buffer: BlockOut Buffer (p/n MB-073) for 1hr at RT. Predicted/Observed size: ~27kDa for GFP.

Western Blot of Goat anti-GFP Antibody with Serums. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: GFP/Human Serum (p/n 000-001-215/D314-05) [0.01/0.02μL] [+]. Lane 3: Human Serum (p/n D314-05) [0.02μL] [-]. Lane 4: GFP/Mouse Serum (p/n 000-001-215/D308-05) [0.01/0.02μL] [+]. Lane 5: Mouse Serum (p/n D308-05) [0.02μL] [-]. Lane 6: GFP/Rat Serum (p/n 000-001-215/D310-05) [0.01/0.02μL] [+]. Lane 7: Rat Serum (p/n D310-05) [0.02μL] [-]. Primary antibody: Anti-GFP antibody at 1.0ug/mL overnight at 2-8°C. Secondary antibody: Donkey Anti-Goat IgG HRP secondary antibody (p/n CUSTmx8) at 1:40,000 for 30 min at RT. Blocking Buffer: BlockOut Buffer (p/n MB-073) for 1hr at RT. Predicted/Observed size: ~27kDa for GFP.

Npas4 expression during neural differentiation of ESCs. (A) RT-PCR analysis was used to determine the temporal expression profile of Npas4 mRNA in relation to various marker genes during N2B27 differentiation of mESCs (n？=？3). Primers to the reference gene β-actin were used as a loading control. The negative control reaction (−) contained water in place of template cDNA. (B) The changes in Npas4 expression during N2B27 differentiation were quantified using qRT-PCR. At each time point Npas4 expression was normalized to β-actin expression and fold changes are relative to Day 0 (undifferentiated mESCs). Mean values and standard deviations of three independent experiments (n？=？3) are displayed. (C)In situ hybridization analysis of Npas4 mRNA expression at Day 4 of N2B27 differentiation of mESCs. Representative images of differentiating colonies from two independent experiments (n？=？2) are shown. Top panel - Npas4 antisense probe; Bottom panel - Npas4 sense probe. Scale bar？=？100？μm. (D) Immunoblotting was used to determine the temporal expression profile of the Npas4 protein during N2B27 differentiation of mESCs. An antibody to the reference protein β-actin was used as a loading control (n？=？3). (E) Immunoblotting was used to determine the temporal expression profile of Sox1 in the 46C cell line (using an antibody to GFP) and βIII tubulin during N2B27 differentiation of mESCs. An antibody to the reference protein β-actin was used as a loading control (n？=？3). (F) RT-PCR analysis of NPAS4 mRNA in relation to various marker genes during Noggin-induced neural differentiation of hESCs (n？=？3). Primers to the reference gene β-ACTIN were used as a loading control. The negative control reaction (−) contained water in place of template cDNA. hESCs, human embryonic stem cells; mESCs, mouse embryonic stem cells; NS, neurospheres. Figure provided by CiteAb. Source: Stem Cell Res Ther, PMID: 24887558.

Tyrosine phosphorylation of HIPK mutants. HeLa cells were transiently transfected with the indicated expression constructs for HIPK1 (A), HIPK2 (B), HIPK3 (C) or HIPK4 (D). Sodium orthovanadate (Na3VO4) was added to every second sample for 1 h before lysis. GFP fusion proteins were immunoprecipitated and analysed by immunodetection with antibodies for pTyr and GFP. The panels are representative of 2？3 independent experiments. Figure provided by CiteAb. Source: Cell Commun Signal, PMID: 25630557.

Tyrosine phosphorylation of HIPK mutants. HeLa cells were transiently transfected with the indicated expression constructs for HIPK1 (A), HIPK2 (B), HIPK3 (C) or HIPK4 (D). Sodium orthovanadate (Na3VO4) was added to every second sample for 1 h before lysis. GFP fusion proteins were immunoprecipitated and analysed by immunodetection with antibodies for pTyr and GFP. The panels are representative of 2？3 independent experiments. Figure provided by CiteAb. Source: Cell Commun Signal, PMID: 25630557.