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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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Simple Western™ analysis of lysates (1 mg/mL) from Raji cells using c-Myc (D84C12) Rabbit mAb #5605. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells treated with UV (100 mJ/cm2; 2H recovery) using c-Jun (60A8) Rabbit mAb #9165. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells serum starved for 24hr and treated with TPA (400nM, 4hr) using c-Raf Antibody #9422. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from various cell lines and tissues using c-Rel (D4Y6M) Rabbit mAb.
Western blot analysis of cell extracts from various cell lines, using Src (32G6) Rabbit mAb.
Western blot analysis of extracts from various cell lines, using c-Abl Antibody.
Western blot analysis of extracts from NCI-H526 cells using c-Kit (D13A2) XP® Rabbit mAb.
Western blot analysis of extracts from 293T, C2C12 and C6 cells using Ras (27H5) Rabbit mAb.
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.
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.
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.
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.
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.
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.
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).
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.
Western blot analysis of extracts from NIH/3T3 and SK-N-MC cells, untreated or UV-treated, using c-Jun (60A8) Rabbit mAb.
Western blot analysis of extracts from various cell lines using c-Myc (D84C12) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human astrocytoma, using c-Jun (60A8) Rabbit mAb.
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).
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using c-Jun (60A8) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using c-Jun (60A8) Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells, using c-Jun (60A8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
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.
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 Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across Dclk1, a known target gene of c-Jun (see additional figure containing ChIP-qPCR data).
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 Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #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).
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.
To Purchase # 9328
Cat. # Size Qty. Price
9328T
1 Kit  (9 x 20 microliters)

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
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).

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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