goat anti cfh  (Quidel)

Journal: Molecular Vision

Article Title: Localization of complement factor H gene expression and protein distribution in the mouse outer retina

doi:

Figure Lengend Snippet: Antibodies, suppliers, and concentrations used for immunohistochemistry and western blots.

Article Snippet: The slides were then incubated overnight at 4 °C in a humidity chamber with 0.8 μg/ml goat anti-mouse Cfh antibody and either 15 μg/ml PNA or 5 μg/ml WGA, both lectins conjugated to fluorescein (Vector Laboratories).

Techniques: Immunohistochemistry, Western Blot, Concentration Assay, Affinity Purification, Recombinant

Journal: Molecular Vision

Article Title: Localization of complement factor H gene expression and protein distribution in the mouse outer retina

doi:

Figure Lengend Snippet: Western blots using different anti-mouse Cfh antibodies. Goat anti-mouse Cfh ( A ) and rabbit anti-mouse Cfh ( B ) antibodies probed against increasing concentrations of human (0.05, 0.1, and 0.2 μg/ml in lanes 2–4, respectively) and mouse (0.05, 0.1, and 0.2 μg/ml in lanes 5–7, respectively) purified complement factor H, and increasing volumes of mouse serum (1, 2.5, 5.0, and 10 μl serum/lane, lanes 8–11, respectively, loaded 40 μl total volume/lane). The goat anti-mouse Cfh antibody recognized human and mouse Cfh, as well as Cfh in mouse serum ( A ), while the rabbit anti-mouse Cfh antibody recognized only mouse Cfh and Cfh in mouse serum ( B ). Nonspecific binding by secondary antibodies was tested by primary antibody deletion (Lane 12). Both antibodies used against serum (1 μl/lane) from 129/Sv Cfh −/− (KO) and their wild-type background strain 129/Sv (WT) recognized Cfh only in serum from the background strain (WT) and C57BL/6 (B6) serum ran as a control ( C ). Note that both antibodies recognized only one major band at approximately 155 kDa (red arrow), the approximate weight of Cfh.

Article Snippet: The slides were then incubated overnight at 4 °C in a humidity chamber with 0.8 μg/ml goat anti-mouse Cfh antibody and either 15 μg/ml PNA or 5 μg/ml WGA, both lectins conjugated to fluorescein (Vector Laboratories).

Techniques: Western Blot, Purification, Binding Assay

Journal: Molecular Vision

Article Title: Localization of complement factor H gene expression and protein distribution in the mouse outer retina

doi:

Figure Lengend Snippet: Immunohistochemistry for Cfh protein expression in the BALB/c, 129/Sv, and 129/Sv Cfh −/− eye. A BALB/c male eye sectioned near the optic nerve labeled for Cfh using two anti-Cfh antibodies ( C , D ), along with its paired negative control ( A , B ). Goat anti-mouse Cfh antibody predominately labeled photoreceptor outer segments (OS) and what appear to be cone cell photoreceptor (arrow heads) inner segments (IS; C ). The rabbit anti-mouse Cfh antibody labeled OS, the apical edge of the retinal pigment epithelium (RPE) cells, and only lightly labeled the RPE cytoplasm ( D ). The 129/Sv background strain eyes and Cfh knockout ( 129/Sv Cfh −/− ) eyes paired with their negative controls ( E , F , I , J ) immunolabeled with goat anti-mouse Cfh antibody ( G , H ) or rabbit anti-mouse Cfh antibody ( K , L ). Cfh labeling was observed in the Cfh knockout eyes ( H , L ) using both antibodies, but the signal was greatly reduced compared to the wild-type (WT) eyes ( G , K ). The residual presence of the signal could be due to cross-reactivity with Cfhr2 protein, since both antibodies recognize Cfh and Cfhr2 proteins. Scale bar=20 μm.

Article Snippet: The slides were then incubated overnight at 4 °C in a humidity chamber with 0.8 μg/ml goat anti-mouse Cfh antibody and either 15 μg/ml PNA or 5 μg/ml WGA, both lectins conjugated to fluorescein (Vector Laboratories).

Techniques: Immunohistochemistry, Expressing, Labeling, Negative Control, Knock-Out, Immunolabeling

Journal: Molecular Vision

Article Title: Localization of complement factor H gene expression and protein distribution in the mouse outer retina

doi:

Figure Lengend Snippet: Cfh localization within rod and cone photoreceptors. A sagittal section from a male BALB/c mouse eye immunolabeled for rhodopsin (green), Cfh (red), and a 4',6-diamidino-2-phenylindole (DAPI) nuclear stain (blue) showed only a localized band of Cfh in the central region of the OS (white arrow), but not throughout the OS. In addition, occasional Cfh labeling of cells believed to be cone cell IS was observed (white arrowheads; A ). A sagittal section from a male BALB/c mouse eye immunolabeled for blue and red/green opsins (white), Cfh (red), and a DAPI nuclear stain (blue) showed a similar band of Cfh in the central region of the OS (green arrow; B ). Cone cells labeled with the opsin antibodies (green arrowheads) exhibit a polarized distribution of blue and red/green opsins (white) and Cfh (red). A coronal section from a male C57BL/6 eye double labeled with fluorescein conjugated WGA (green), which labels the matrix domains surrounding rod photoreceptors, and Cfh (red) showing Cfh within rod photoreceptor OS ( C ). The delineated area of interest is magnified and shown on the right. Cfh is clearly localized within the rods (arrows; D ). A coronal section from a male C57BL/6 eye labeled with fluorescein conjugated PNA (green), which binds cone matrix domains, and Cfh (red) showing Cfh is clearly localized within the cone cells (arrows; E ). The delineated area of interest is magnified and shown on the right ( F ). IS=photoreceptor inner segments; OS=outer segments; RPE=retinal pigment epithelium; WGA=wheat germ agglutinin; PNA=peanut agglutinin. Scale bar=20 μm.

Article Snippet: The slides were then incubated overnight at 4 °C in a humidity chamber with 0.8 μg/ml goat anti-mouse Cfh antibody and either 15 μg/ml PNA or 5 μg/ml WGA, both lectins conjugated to fluorescein (Vector Laboratories).

Techniques: Immunolabeling, Staining, Labeling