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8333
Fos Family Antibody Sampler Kit

Fos Family Antibody Sampler Kit #8333

Western Blotting Image 1

Western blot analysis of extracts from HeLa, RAW, and H-4-IIE cells serum-starved overnight and TPA-stimulated for 4 hours, using c-Fos Antibody.

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Western Blotting Image 2

Western blot analysis of extracts from HeLa cells, serum-starved overnight and then either untreated or stimulated for 4 hours with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174, using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb (upper) and c-Fos (9F6) Rabbit mAb #2250 (lower). Antibody phospho-specificity is shown by treating lysates with λ-phosphatase.

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Chromatin IP-seq Image 3

Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with β-NGF #5221 (50 ng/ml) for 2 hr and FosB (5G4) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across Ccrn4l, a known target gene of FosB (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 4

Western blot analysis of extracts from HeLa cells serum-starved overnight and TPA-stimulated for 4 hours, or NIH/3T3 cells and C6 cells serum-starved overnight and serum-stimulated for 4 hours, using FosB (5G4) Rabbit mAb.

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Western Blotting Image 5

Western blot analysis of extracts from serum-starved HeLa cells, untreated or stimulated with TPA #4174 for 4 hours, using FRA1 (D80B4) Rabbit mAb (upper) and β-Actin Antibody #4967 (lower).

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Western Blotting Image 6

Western blot analysis of extracts from HeLa cells, serum-starved overnight, and either left untreated or treated with TPA #4174 for 4 hours, using Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb #5841 (upper) and FRA1 (D80B4) Rabbit mAb #5281 (lower). Antibody phospho-specificity is shown by treating lysates with λ phosphatase.

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Western Blotting Image 7

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Flow Cytometry Image 8

Flow cytometric analysis of HeLa cells, untreated (blue) or TPA-treated (green), using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb.

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IHC-P (paraffin) Image 9

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using FosB (5G4) Rabbit mAb.

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Chromatin IP Image 10

Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with β-NGF #5221 (50 ng/ml) for 2 hr and either FosB (5G4) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR SimpleChIP® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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Chromatin IP Image 11

Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and either Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR SimpleChIP® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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IF-IC Image 12

Confocal immunofluorescent analysis of HeLa cells, serum-starved (left), TPA-treated (middle) or treated with TPA and λ-phosphatase (right), using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using FosB (5G4) Rabbit mAb in the presence of control peptide (left) or FosB Blocking Peptide #1042 (right).

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Chromatin IP Image 14

Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with Human β-Nerve Gowth Factor (hβ-NGF) #5221 (50ng/ml) for 2h and either Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded HeLa cells control (left) or PMA-treated (right), using FosB (5G4) Rabbit mAb.

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Flow Cytometry Image 16

Flow cytometric analysis of HeLa cells, untreated (blue) or TPA treated (green), using FosB (5G4) Rabbit mAb compared to a nonspecific negative control antibody (red).

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IF-IC Image 17

Confocal immunofluorescent analysis of HeLa cells either serum-starved (left) or TPA-treated (right) and labeled with FosB (5G4) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
c-Fos Antibody 4384 20 µl
  • WB
H M R 62 Rabbit 
Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb 5348 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M R 62 Rabbit IgG
FosB (5G4) Rabbit mAb 2251 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R 38 FosB2 48 FosB Rabbit IgG
FRA1 (D80B4) Rabbit mAb 5281 20 µl
  • WB
  • IP
H 40 Rabbit IgG
Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb 5841 20 µl
  • WB
  • ChIP
H M R 40 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Fos Family Antibody Sampler Kit provides an economical means to evaluate the Fos family of transcription factors. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Each antibody in the Fos Family Antibody Sampler Kit recognizes endogenous levels of the specific target protein. FosB (5G4) Rabbit mAb detects both FosB and FosB2 isoforms. Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb recognizes endogenous levels of FRA1 protein only when phosphorylated at Ser265. This antibody may also cross-react with phospho-FRA2, but does not cross-react with phospho-c-Fos or phospho-FosB.

Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser32 of human c-Fos, Ser265 of human FRA1 protein or with a synthetic peptide derived from human FosB, or FRA1.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids near the carboxy-terminus of human c-Fos protein. Antibodies are purified by protein A and peptide affinity chromatography.

The Fos family of nuclear oncogenes includes c-Fos, FosB, Fos-related antigen 1 (FRA1), and Fos-related antigen 2 (FRA2) (1). While most Fos proteins exist as a single isoform, the FosB protein exists as two isoforms: full-length FosB and a shorter form, FosB2 (Delta FosB), which lacks the carboxy-terminal 101 amino acids (1-3). The expression of Fos proteins is rapidly and transiently induced by a variety of extracellular stimuli including growth factors, cytokines, neurotransmitters, polypeptide hormones, and stress. Fos proteins dimerize with Jun proteins (c-Jun, JunB, and JunD) to form Activator Protein-1 (AP-1), a transcription factor that binds to TRE/AP-1 elements and activates transcription. Fos and Jun proteins contain the leucine-zipper motif that mediates dimerization and an adjacent basic domain that binds to DNA. The various Fos/Jun heterodimers differ in their ability to transactivate AP-1 dependent genes. In addition to increased expression, phosphorylation of Fos proteins by Erk kinases in response to extracellular stimuli may further increase transcriptional activity (4-6). Phosphorylation of c-Fos at Ser32 and Thr232 by Erk5 increases protein stability and nuclear localization (5). Phosphorylation of FRA1 at Ser252 and Ser265 by Erk1/2 increases protein stability and leads to overexpression of FRA1 in cancer cells (6). Following growth factor stimulation, expression of FosB and c-Fos in quiescent fibroblasts is immediate, but very short-lived, with protein levels dissipating after several hours (7). FRA1 and FRA2 expression persists longer, and appreciable levels can be detected in asynchronously growing cells (8). Deregulated expression of c-Fos, FosB, or FRA2 can result in neoplastic cellular transformation; however, Delta FosB lacks the ability to transform cells (2,3).

  1. Tulchinsky, E. (2000) Histol Histopathol 15, 921-8.
  2. Dobrazanski, P. et al. (1991) Mol Cell Biol 11, 5470-8.
  3. Nakabeppu, Y. and Nathans, D. (1991) Cell 64, 751-9.
  4. Rosenberger, S.F. et al. (1999) J Biol Chem 274, 1124-30.
  5. Sasaki, T. et al. (2006) Mol Cell 24, 63-75.
  6. Basbous, J. et al. (2007) Mol Cell Biol 27, 3936-50.
  7. Kovary, K. and Bravo, R. (1991) Mol Cell Biol 11, 2451-9.
  8. Kovary, K. and Bravo, R. (1992) Mol Cell Biol 12, 5015-23.
Entrez-Gene Id
2353 , 2354 , 8061
Swiss-Prot Acc.
P01100 , P53539 , P15407
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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