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8333
Fos Family Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Fos Family Antibody Sampler Kit #8333

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Fos Family Antibody Sampler Kit: 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.

Fos Family Antibody Sampler Kit: Image 2

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 Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across Fbxl19 gene. For additional ChIP-seq tracks, please download the product data sheet.

Fos Family Antibody Sampler Kit: Image 3

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).

Fos Family Antibody Sampler Kit: Image 4

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.

Fos Family Antibody Sampler Kit: Image 5

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.

Fos Family Antibody Sampler Kit: Image 6

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.

Fos Family Antibody Sampler Kit: Image 7

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.

Fos Family Antibody Sampler Kit: Image 8

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).

Fos Family Antibody Sampler Kit: Image 9

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.

Fos Family Antibody Sampler Kit: Image 10

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

Fos Family Antibody Sampler Kit: Image 11

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).

Fos Family Antibody Sampler Kit: Image 12

Confocal immunofluorescent analysis of mouse cerebral cortex both untreated (left) and lambda-phosphatase treated (right) using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb #5348 (green) and Neurofilament-L (DA2) Mouse mAb #2835 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

Fos Family Antibody Sampler Kit: Image 13

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

Fos Family Antibody Sampler Kit: Image 14

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).

Fos Family Antibody Sampler Kit: Image 15

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).

Fos Family Antibody Sampler Kit: Image 16

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.

Fos Family Antibody Sampler Kit: Image 17

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

Fos Family Antibody Sampler Kit: Image 18

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.

To Purchase # 8333T
Product # Size Price
8333T
1 Kit  (5 x 20 µl) $ 391

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 

Product Description

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.

Specificity / Sensitivity

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.

Source / Purification

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.

Background

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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