Product overview
| Name | Anti-c-Fos antibody ValidAb™ |
| Host | Mouse |
| Clonality | Monoclonal |
| Target | c-Fos |
| Description | Antibody to c-Fos - an immediate early gene used as a marker of neuronal activity. Part of the ValidAb™ range of highly validated, data-rich antibodies. |
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Validation data
Figure 1. c-Fos expression in the dentate gyrus of TRAP2;Ai14 mice.
HB8006 reveals c-Fos positive neurons activated during re-exposure to a learning event alongside tdTomato expression in cells activated during the initial learning event. Method: TRAP2;Ai14 mice were administered tamoxifen and underwent a learning experience (driving expression of tdTomato in activated neurones). Seven days later animals were re-exposed to the stimulus then trans-cardiac perfused 90 minutes after behaviour with 0.1M PB (pH7.4) followed by 4% PFA. Brains were extracted and then post-fixed in 4% PFA for 2 hours followed by incubation in 30% sucrose until the brain sank. Coronal sections were cut on a cryostat and stored in cryopreserve at -20°C before further processing. Sections were washed in PBS before being incubated for 30 minutes in sodium citrate buffer (pH 6.0) at 80°C. After allowing sections to return to ambient temperature they were washed in PBST (PBS, 0.2% Triton X-100) before being incubated in mouse on mouse blocking buffer for 1 hour (2 drops per 2.5ml PBST, Vector laboratories, MKB-2213-1). Sections were then incubated in 5% goat serum, 2.5% BSA (blocking buffer) for 1 hour before being incubated for 48hrs at 4°C in HB8006 (1:1000 dilution in blocking buffer, 1µg/ml). Following washing, sections were then incubated in secondary antibody (Goat anti-mouse Alexa Fluor 488 conjugated, 1:500 dilution in blocking buffer, Invitrogen, A-11029) for 24hrs at 4°C. Following washing, sections were mounted using Vectashield Hardset with DAPI (H-1500) and imaged using a 100x objective using a epifluorescence microscope. Data was kindly provided by Gareth Barker at the University of Bristol.
Figure 2. c-Fos expression in the cortex of TRAP2;Ai14 mice.
HB8006 reveals c-Fos positive neurons activated during re-exposure to a learning event alongside tdTomato expression in cells activated during the initial learning event. Method: TRAP2;Ai14 mice were administered tamoxifen and underwent a learning experience (driving expression of tdTomato in activated neurones). Seven days later animals were re-exposed to the stimulus then trans-cardiac perfused 90 minutes after behaviour with 0.1M PB (pH7.4) followed by 4% PFA. Brains were extracted and then post-fixed in 4% PFA for 2 hours followed by incubation in 30% sucrose until the brain sank. Coronal sections were cut on a cryostat and stored in cryopreserve at -20°C before further processing. Sections were washed in PBS before being incubated for 30 minutes in sodium citrate buffer (pH 6.0) at 80°C. After allowing sections to return to ambient temperature they were washed in PBST (PBS, 0.2% Triton X-100) before being incubated in mouse on mouse blocking buffer for 1 hour (2 drops per 2.5ml PBST, Vector laboratories, MKB-2213-1). Sections were then incubated in 5% goat serum, 2.5% BSA (blocking buffer) for 1 hour before being incubated for 48hrs at 4°C in HB8006 (1:1000 dilution in blocking buffer, 1µg/ml). Following washing, sections were then incubated in secondary antibody (Goat anti-mouse Alexa Fluor 488 conjugated, 1:500 dilution in blocking buffer, Invitrogen, A-11029) for 24hrs at 4°C. Following washing, sections were mounted using Vectashield Hardset with DAPI (H-1500) and imaged using a 100x objective using a epifluorescence microscope. Data was kindly provided by Gareth Barker at the University of Bristol.
Figure 3. c-Fos expression in mouse dentate gyrus.
HB8006 reveals c-Fos positive neurons activated during re-exposure to a learning event. Method: TRAP2;Ai14 mice underwent an initial learning experience followed by re-exposure 7 days later. 90 minutes after behaviour, animals were trans-cardiac perfused with 0.1M PB (pH7.4) followed by 4% PFA. Brains were extracted and then post-fixed in 4% PFA for 2 hours followed by incubation in 30% sucrose until the brain sank. Coronal sections were cut on a cryostat and stored in cryopreserve at -20°C before further processing. Sections were washed in PBS before being incubated for 30 minutes in sodium citrate buffer (pH 6.0) at 80°C. After allowing sections to return to ambient temperature they were washed in PBST (PBS, 0.2% Triton X-100) before being incubated in mouse on mouse blocking buffer for 1 hour (2 drops per 2.5ml PBST, Vector laboratories, MKB-2213-1). Sections were then incubated in 5% goat serum, 2.5% BSA (blocking buffer) for 1 hour before being incubated for 48hrs at 4°C in HB8006 (1:1000 dilution in blocking buffer, 1µg/ml). Following washing, sections were then incubated in secondary antibody (Goat anti-mouse Alexa Fluor 488 conjugated, 1:500 dilution in blocking buffer, Invitrogen, A-11029) for 24hrs at 4°C. Following washing, sections were mounted using Vectashield Hardset with DAPI (H-1500) and imaged using a 100x objective using a epifluorescence microscope. Data was kindly provided by Gareth Barker at the University of Bristol.
Figure 1. c-Fos expression in the dentate gyrus of TRAP2;Ai14 mice.
HB8006 reveals c-Fos positive neurons activated during re-exposure to a learning event alongside tdTomato expression in cells activated during the initial learning event. Method: TRAP2;Ai14 mice were administered tamoxifen and underwent a learning experience (driving expression of tdTomato in activated neurones). Seven days later animals were re-exposed to the stimulus then trans-cardiac perfused 90 minutes after behaviour with 0.1M PB (pH7.4) followed by 4% PFA. Brains were extracted and then post-fixed in 4% PFA for 2 hours followed by incubation in 30% sucrose until the brain sank. Coronal sections were cut on a cryostat and stored in cryopreserve at -20°C before further processing. Sections were washed in PBS before being incubated for 30 minutes in sodium citrate buffer (pH 6.0) at 80°C. After allowing sections to return to ambient temperature they were washed in PBST (PBS, 0.2% Triton X-100) before being incubated in mouse on mouse blocking buffer for 1 hour (2 drops per 2.5ml PBST, Vector laboratories, MKB-2213-1). Sections were then incubated in 5% goat serum, 2.5% BSA (blocking buffer) for 1 hour before being incubated for 48hrs at 4°C in HB8006 (1:1000 dilution in blocking buffer, 1µg/ml). Following washing, sections were then incubated in secondary antibody (Goat anti-mouse Alexa Fluor 488 conjugated, 1:500 dilution in blocking buffer, Invitrogen, A-11029) for 24hrs at 4°C. Following washing, sections were mounted using Vectashield Hardset with DAPI (H-1500) and imaged using a 100x objective using a epifluorescence microscope. Data was kindly provided by Gareth Barker at the University of Bristol.
Figure 2. c-Fos expression in the cortex of TRAP2;Ai14 mice.
HB8006 reveals c-Fos positive neurons activated during re-exposure to a learning event alongside tdTomato expression in cells activated during the initial learning event. Method: TRAP2;Ai14 mice were administered tamoxifen and underwent a learning experience (driving expression of tdTomato in activated neurones). Seven days later animals were re-exposed to the stimulus then trans-cardiac perfused 90 minutes after behaviour with 0.1M PB (pH7.4) followed by 4% PFA. Brains were extracted and then post-fixed in 4% PFA for 2 hours followed by incubation in 30% sucrose until the brain sank. Coronal sections were cut on a cryostat and stored in cryopreserve at -20°C before further processing. Sections were washed in PBS before being incubated for 30 minutes in sodium citrate buffer (pH 6.0) at 80°C. After allowing sections to return to ambient temperature they were washed in PBST (PBS, 0.2% Triton X-100) before being incubated in mouse on mouse blocking buffer for 1 hour (2 drops per 2.5ml PBST, Vector laboratories, MKB-2213-1). Sections were then incubated in 5% goat serum, 2.5% BSA (blocking buffer) for 1 hour before being incubated for 48hrs at 4°C in HB8006 (1:1000 dilution in blocking buffer, 1µg/ml). Following washing, sections were then incubated in secondary antibody (Goat anti-mouse Alexa Fluor 488 conjugated, 1:500 dilution in blocking buffer, Invitrogen, A-11029) for 24hrs at 4°C. Following washing, sections were mounted using Vectashield Hardset with DAPI (H-1500) and imaged using a 100x objective using a epifluorescence microscope. Data was kindly provided by Gareth Barker at the University of Bristol.
Figure 3. c-Fos expression in mouse dentate gyrus.
HB8006 reveals c-Fos positive neurons activated during re-exposure to a learning event. Method: TRAP2;Ai14 mice underwent an initial learning experience followed by re-exposure 7 days later. 90 minutes after behaviour, animals were trans-cardiac perfused with 0.1M PB (pH7.4) followed by 4% PFA. Brains were extracted and then post-fixed in 4% PFA for 2 hours followed by incubation in 30% sucrose until the brain sank. Coronal sections were cut on a cryostat and stored in cryopreserve at -20°C before further processing. Sections were washed in PBS before being incubated for 30 minutes in sodium citrate buffer (pH 6.0) at 80°C. After allowing sections to return to ambient temperature they were washed in PBST (PBS, 0.2% Triton X-100) before being incubated in mouse on mouse blocking buffer for 1 hour (2 drops per 2.5ml PBST, Vector laboratories, MKB-2213-1). Sections were then incubated in 5% goat serum, 2.5% BSA (blocking buffer) for 1 hour before being incubated for 48hrs at 4°C in HB8006 (1:1000 dilution in blocking buffer, 1µg/ml). Following washing, sections were then incubated in secondary antibody (Goat anti-mouse Alexa Fluor 488 conjugated, 1:500 dilution in blocking buffer, Invitrogen, A-11029) for 24hrs at 4°C. Following washing, sections were mounted using Vectashield Hardset with DAPI (H-1500) and imaged using a 100x objective using a epifluorescence microscope. Data was kindly provided by Gareth Barker at the University of Bristol.
Product information
| Immunogen | Full length recombinant human c-Fos expressed in and purified from E. coli. |
| Clone number | 2H2 |
| Isotype | IgG1 |
| Purification | Protein G affinity chromatography |
| Concentration | 1 mg/ml |
| Formulation | 50% PBS, 50% glycerol with 5mM sodium azide |
| Predicted species reactivity | Mouse, Rat, Human |
| Tested species reactivity | Mouse |
Tested applications
| Applications | IHC(IF) |
| IHC(IF) optimal concentration | 1μg/ml (1:1000 dilution) as tested in 4% PFA fixed mouse brain tissue |
| Product specific protocols |
|
| Positive control | c-Fos is expressed at high levels in many cell lines (e.g. HEK293 or HeLa) when serum starved cells are stimulated with serum. |
| Negative control | Serum starved cell lines (e.g. HEK293 or HeLa) express very low levels of c-Fos. |
| Open data link | Please follow this link to the OSF |
Target information
| Other names |
|
| UniProt ID | P01100 |
| Structure image |  |
| Gene name | FOS |
| NCBI full gene name | Fos proto-oncogene, AP-1 transcription factor subunit |
| Entrez gene ID |
| Amino acids | 380aa (40.7kDa) |
| Isoforms | c-Fos has three key isoforms:
|
| Expression | Expressed widely across multiple tissues at low levels. Expression is inducible by a range of factors including cellular activity, growth factors, cytokines and tumour promoters. c-Fos is expressed in neurones where its expression is induced by activity. |
| Subcellular expression | c-Fos expression is localised to the nucleus |
| Processing | None |
| Post translational modifications | Subject to phosphorylation on multiple residues and also possesses multiple SUMO2 binding sites. |
| Homology (compared to human) | Rat and mouse show a 94.2% and 93.7% identity to human c-Fos in a BLAST search |
| Similar proteins | The following proteins were identified as being similar to c-Fos in a BLAST search:
|
Storage & Handling
| Storage instructions | -20°C |
| Important | This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use |
Technical guides
Western Blot Protocol (1 MB) Immunocytochemistry Protocol (1.2 MB) Immunohistochemistry Protocol (1.6 MB) References for Anti-c-Fos antibody ValidAb™
References are publications that support the biological activity of the product
-
Adult social isolation leads to anxiety and spatial memory impairment: Brain activity pattern of COx and c-Fos.
Zorzo C et al (2019) Behavioural brain research 365 : 170-177 -
Decreased food anticipatory activity of obese mice relates to hypothalamic c-Fos expression.
Luna-Illades C et al (2017) Physiology & behavior 179 : 9-15 -
c-FOS expression after hippocampal deep brain stimulation in normal rats.
da Silva JC et al (2014) Neuromodulation : journal of the International Neuromodulation Society 17 : 213-7; discussion 216-7 -
Activation of c-fos in the brain.
Herrera DG et al (1996) Progress in neurobiology 50 : 83-107 -
Expression of c-fos-like protein as a marker for neuronal activity following noxious stimulation in the rat.
Bullitt E (1990) The Journal of comparative neurology 296 : 517-30
Publications
These publications cite the use of Anti-c-Fos antibody ValidAb™ purchased from Hello Bio:
-
Adrenergic C1 neurons enhance anxiety via projections to PAG
Fernandez-Pena et al (2024) bioRxiv
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