Rabbit anti-C-FOS Antibody

Name

Rabbit anti-C-FOS Antibody

中文名字

兔抗C-FOS抗体

Description

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.

Clonality

Polyclonal

Isotype

IgG

Host

Rabbit

Quantity/Volume

50ul/100 µL

State

Liquid Whole Serum/antibody

Reacts With

Mouse, Rat

Preabsorption Control

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Alternate Names

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

RRID

AB_572267

Immunogen

This information is proprietary to Mabioway

Gene Symbol

FOS

Entrez Gene ID

Entrez Gene:  2353  Human
Entrez Gene: 14281  Mouse
Entrez Gene: 314322  Rat

NCBI Gene Aliases

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Sequence

>sp|P01100|FOS_HUMAN Protein c-Fos OS=Homo sapiens

APPLICATION

Quality Control

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 significant labeling of rat paraventricular and supraoptic nuclei using indirect immunofluorescent and biotin/avidin-HRP techniques. The antibody was also validated by challenge with the selective 5HT2a agonist TCB2 (10 mg/kg ip) with results showing massive numbers of cortical pyramidal cells of the TCB2 treated rats, consistent with the distribution of 5HT2a receptors. No labeling was seen in negative control rats. 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.

Tissue

Rat brain hypothalamus (paraventricular and supraoptic nuclei) and cortex.

Perfusion Fixation

• Fixative: 4% paraformaldehyde-0.05% glutaraldehye in 0.1M Phosphate buffer, pH 7.4; 500 mL over 20– 30 min.
• Post Fixation: 1.5 hour at 4°C in 4% paraformaldehyde-0.05% glutaraldehye in 0.1M phosphate buffer, pH 7.4.

Absorption Control

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Sections

50 µm vibratome

Tissue Incubation

18–24 hours at 2°–8°C

Detection System

Use IF or Bn/Av-HRP reagents at dilutions recommended by the manufacturer.

Suggested Dilution

1/4,000–1/6,000 in PBS/0.3% Triton X-100 – Bn/Av-HRP immunohistochemistry

NOTES

Special Instructions

It is recommended that the researcher perform a primary antibody dilution series using our dilution recommendations as a guideline. Note that a change in the fixation or buffering system from our protocol may change the configuration of the protein which could alter the reactivity with the tissue tested.

Concentration

Not applicable. Antibody concentration is only relevant for purified antibodies.

Storage

After reconstitution, use immediately or refrigerate at 2º–8ºC up to 2 days. For long term storage, aliquot and freeze at -15°C or lower. Avoid repeated freeze/thaw cycles.