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9328
c-Oncogene Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

c-Oncogene Antibody Sampler Kit #9328

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other Image 1 - c-Oncogene Antibody Sampler Kit

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.

other Image 2 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from various cell lines, using c-Abl Antibody.

other Image 3 - c-Oncogene Antibody Sampler Kit

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 c-Jun (60A8) 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 Dclk1, a known target gene of c-Jun (see additional figure containing ChIP-qPCR data).

other Image 4 - c-Oncogene Antibody Sampler Kit

Confocal immunofluorescent analysis of NCI-H526 (left) and Jurkat (right) cells using c-Kit (D13A2) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

other Image 5 - c-Oncogene Antibody Sampler Kit

Flow cytometric analysis of Raji cells using c-Myc (D84C12) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

other Image 6 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from HeLa, C2C12 or PC12 cells, untreated or TPA-treated (200 nM for 30 minutes), using c-Raf Antibody #9422.

other Image 7 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from 293T, C2C12 and C6 cells using Ras (27H5) Rabbit mAb.

other Image 8 - c-Oncogene Antibody Sampler Kit

Western blot analysis of cell extracts from various cell lines, using Src (32G6) Rabbit mAb.

other Image 9 - c-Oncogene Antibody Sampler Kit

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.

other Image 10 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from various cell lines and tissues using c-Rel (D4Y6M) Rabbit mAb.

other Image 11 - c-Oncogene Antibody Sampler Kit

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 c-Jun (60A8) 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 chromosome 2 (upper), including Dclk1 (lower), a known target gene of c-Jun (see additional figure containing ChIP-qPCR data).

other Image 12 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from NCI-H526 cells using c-Kit (D13A2) XP® Rabbit mAb.

other Image 13 - c-Oncogene Antibody Sampler Kit

Confocal immunofluorescent analysis of HeLa cells, mock-transfected (left) or transfected with SignalSilence® c-Myc siRNA I #6341 (right), using c-Myc (D84C12) Rabbit mAb (green). Actin filaments have been labeled wth DY-554 phalloidin (red).

other Image 14 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from Neuro-2a cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® c-Rel siRNA I (Mouse Specific) #13058 (+) or SignalSilence® c-Rel siRNA II (Mouse Specific) #13170 (+), using c-Rel (D4Y6M) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The c-Rel (D4Y6M) Rabbit mAb confirms silencing of c-Rel expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

other Image 15 - c-Oncogene Antibody Sampler Kit

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 of c-Jun (60A8) 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.

other Image 16 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from control HEK293 cells (lane 1) or c-Myc knockout HEK293 cells (lane 2) using c-Myc (D84C12) Rabbit mAb Antibody, #5605 (upper) or β-actin (13E5) Rabbit mAb, #4970 (lower). The absence of signal in the c-Myc knockout HEK293 cells confirms specificity of the antibody for c-Myc.

other Image 17 - c-Oncogene Antibody Sampler Kit

Flow cytometric analysis of Jurkat cells using c-Jun (60A8) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

other Image 18 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from HeLa cells, mock transfected or transfected with SignalSilence® c-Myc siRNA I #6341, using c-Myc (D84C12) Rabbit mAb.

other Image 19 - c-Oncogene Antibody Sampler Kit

Confocal immunofluorescent analysis of HeLa cells, using c-Jun (60A8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

other Image 20 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from various cell lines using c-Myc (D84C12) Rabbit mAb.

other Image 21 - c-Oncogene Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human astrocytoma, using c-Jun (60A8) Rabbit mAb.

other Image 22 - c-Oncogene Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using c-Jun (60A8) Rabbit mAb.

other Image 23 - c-Oncogene Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using c-Jun (60A8) Rabbit mAb.

other Image 24 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from control HeLa cells (lane 1) or c-Jun knockout HeLa cells (lane 2) using c-Jun (60A8) Rabbit mAb #9165. The absence of signal in the c-Jun knockout HeLa cells confirms specificity of the antibody for c-Jun.

other Image 25 - c-Oncogene Antibody Sampler Kit

Western blot analysis of extracts from NIH/3T3 and SK-N-MC cells, untreated or UV-treated, using c-Jun (60A8) Rabbit mAb.

To Purchase # 9328T
Product # Size Price
9328T
1 Kit  (9 x 20 µl) $ 609

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
c-Fos Antibody 4384 20 µl
  • WB
H M R 62 Rabbit 
c-Abl Antibody 2862 20 µl
  • WB
  • IP
H M R 135 (c-Abl); 210 (Bcr-Abl) Rabbit 
c-Jun (60A8) Rabbit mAb 9165 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 43, 48 Rabbit IgG
c-Kit (D13A2) XP® Rabbit mAb 3074 20 µl
  • WB
  • IP
  • IF
H M 120 and 145 Rabbit 
c-Myc (D84C12) Rabbit mAb 5605 20 µl
  • WB
  • IF
  • F
H M R 57-65 Rabbit IgG
c-Raf Antibody 9422 20 µl
  • WB
H M R Mk 65 to 75 Rabbit 
Ras (27H5) Rabbit mAb 3339 20 µl
  • WB
H M R Mk Dm 21 Rabbit IgG
Src (32G6) Rabbit mAb 2123 20 µl
  • WB
  • IP
H M R Mk 60 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
c-Rel (D4Y6M) Rabbit mAb 12707 20 µl
  • WB
H M R 68-78 Rabbit IgG

Product Description

The c-Oncogene Antibody Sampler Kit provides an economical means of evaluating total levels of various oncogenic proteins. The kit contains enough primary and secondary antibodies to perform two Western blot experiments.

Specificity / Sensitivity

Unless otherwise indicated, each antibody in the c-Oncogene Antibody Sampler Kit detects endogenous levels of total target protein and does not cross-react with related proteins. c-Jun (60A8) Rabbit mAb detects endogenous levels of total c-Jun protein, regardless of phosphorylation state. Ras (27H5) Rabbit mAb detects endogenous levels of total K-Ras, H-Ras and N-Ras proteins. Src (32G6) Rabbit mAb detects endogenous levels of Src proteins and does not cross-react with other Src family members. The c-Myc (D84C12) Rabbit mAb detects endogenous levels of total c-Myc protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues near the carboxy terminus of human c-Rel, residues near the carboxy-terminus of human c-Fos, and corresponding to the sequence close to the carboxy-terminus of human c-Abl. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a recombinant fusion protein corresponding to residues 1-110 of human Src, residues near the amino terminus of human K-Ras, from the amino-terminal sequence of human c-Jun, residues near the amino terminus of c-Myc and corresponding to the residues surrounding Tyr703 of human c-Kit.

Background

The regulation of cell growth, differentiation and programmed death is coordinated by several sets of proteins that comprise essential signal transduction pathways. Many of these key regulatory proteins are encoded by proto-oncogenes, which can be activated (altered) to change the typical cell program to one of abnormal cell growth and unregulated development. Proteins encoded by proto-oncogenes include growth factors and other ligands, receptor proteins, tyrosine kinases, various regulatory proteins (i.e. GTPases) and transcription factors. Together these proteins comprise the basic elements of cell signaling pathways; altered expression or mutation of one or more of these components can lead to oncogenic growth (reviewed in 1).

Non-receptor (i.e. cytoplasmic, nuclear) tyrosine kinases such as c-Abl and Src play key roles in the regulation of cell proliferation, differentiation, apoptosis, cell adhesion and stress responses (2,3). Alteration of the corresponding c-Abl and Src proto-oncogenes is associated with oncogenesis; Abl1-BCR gene translocations result in chronic myelogenous leukemia (CML) while constitutively active Src is seen in some patients with colon cancer and altered Src expression is seen in a wide array of cancers (2,4). Regulation of Raf tyrosine kinase by Ras GTPase controls downstream kinases in the MEK/MAPK signaling pathway (5). Activation of the Ras and Raf proto-oncogenes are common in human cancers and both proteins are seen as potential therapeutic targets (6). The receptor tyrosine kinase c-Kit plays a critical role in activation and growth of hematopoietic stem cells (7); mutations that inhibit c-Kit kinase activity are associated with a variety of developmental disorders while mutations producing constitutively active c-Kit can result in mastocytosis and gastrointestinal stromal tumors (8). The alteration of key transcription factors such as c-Fos, c-Jun, c-Myc and c-Rel that are normally responsible for regulating cell and tissue growth, differentiation and the inflammation/immune response, can also result in unregulated, oncogenic cell growth (9-12).

  1. Croce, C.M. (2008) N Engl J Med 358, 502-11.
  2. Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609.
  3. Avruch, J. et al. (1994) Trends Biochem Sci 19, 279-83.
  4. Wang, J.Y. (2000) Oncogene 19, 5643-50.
  5. Dehm, S.M. and Bonham, K. (2004) Biochem Cell Biol 82, 263-74.
  6. Stites, E.C. et al. (2007) Science 318, 463-7.
  7. Gommerman, J.L. et al. (1997) J Biol Chem 272, 30519-25.
  8. Nocka, K. et al. (1990) EMBO J 9, 1805-13.
  9. Milde-Langosch, K. (2005) Eur J Cancer 41, 2449-61.
  10. Shaulian, E. and Karin, M. (2002) Nat Cell Biol 4, E131-6.
  11. Yokota, J. et al. (1986) Science 231, 261-5.
  12. Rayet, B. and Gélinas, C. (1999) Oncogene 18, 6938-47.
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