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.
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
Due to the low stability of the c-Fos protein (around a 1hr half life) we recommend perfusion fixation for animal tissues to ensure rapid preservation of protein integrity. A protocol for this is available in Gage et al., 2012.
This product has only been validated in IHC(IF) using sodium citrate antigen retrieval (pH6.0, 80°C for 30 minutes) and therefore it is highly recommended that this method is used in all IHC(IF) experiments using HB8006.
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.
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:
FosB - 73.9% identity
FRA-1 - 52.7% identity
FRA-2 - 42.9% identity
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
Will my c-Fos antibody work against species that have not been listed on the datasheet?
A species not being listed doesn’t mean that the c-Fos antibody won’t work, just that we haven’t tested it. If you test one of our antibodies in a new species please let us know (positive or negative)!
What protocols are available for use with this c-Fos antibody?
We have made a comprehensive collection of protocols that we have used in our experiments to validate this c-Fos antibody.
What guarantee do you have that my c-Fos antibody will perform as expected?
We guarantee that your c-Fos antibody will work for the applications and species we list on the datasheet. If the antibody fails to perform as expected then we are happy to offer a 100% refund guarantee. For more details please see our guarantee policy.
What mounting media do you recommend to use with this antibody?
We recommend using one of our high performance mounting medias, supplied as either hardset or aqeous with a range of counterstains:
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.
Western Blot Protocol (1 MB) 
