For induction of c-fos protein activity rats were injected with 1.0 ml of 1.5 M NaCl per 100 grams of body weight. Negative control rats were injected with the same volume of normal saline. The ImmunoStar c-fos antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat paraventricular nucleus and supraoptic nucleus using indirect immunofluorescent and biotin/avidin-HRP techniques. No labeling was seen in negative control rats. Recommended primary dilutions for these methods are 1/4000-1/6000 in PBS/0.3% Triton X-100 – FITC Technique and 1/4000-1/6000 in PBS/0.3% Triton X-100 – biotin/avidin-HRP Technique.

Injection of intraperitoneal hypertonic saline to provoke fos expression is most usually used to set ICC staining parameters.  It is critical that tissue is harvested 60-90 minutes post injection for this and any other stress that is used.  Inject 1 ml/100 g body weight 1.5 M NaCl intraperitoneally.  Harvest brains 90 minutes post injection.  Time is critical.

Specificity of the antiserum was demonstrated by blockage of staining in experimental rats by omission of c-fos antibody or by substitution of antibody pre-incubated with synthetic peptide or the conjugate. Immunoblot analysis of mediobasal hypothalamus showed a single band of approximately 55-60 kD.

照片描述: IHC image of neurons staining for c-Fos in the rat supraoptic nucleus. The tissue was fixed with 4% formaldehyde in 0.1 M phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:5000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen. The section was then mounted on slides with permount.

宿主: Rabbit

规格/体积: 100 µL

性状: Lyophilized Whole Serum

反应性: Mouse, Rat

别名:  Proto-oncogene c-Fos; Cellular oncogene; G0/G1 switch regulatory protein 7; Proto-oncogene protein; G0S7; p55; AP-1; FBJ murine osteosarcoma viral oncogene homolog, anti-CFOS

Gene Symbol:  FOS

RRID:  AB_572267