Interested in promotions? | Click here >>
26717
Myc Family Profiling Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Myc Family Profiling Antibody Sampler Kit #26717

Reviews ()
Citations (0)
Myc Family Profiling Antibody Sampler Kit: Image 1
Western blot analysis of extracts from KARPAS-299, cells untreated (-) or treated with MG-132 #2194 (10 μM, overnight; +) using Phospho-c-Myc (Ser62) (E1J4K) Rabbit mAb. The phospho-specificity of the antibody was demonstrated by peptide competition using phosphopeptides containing Ser62, Thr58/Ser62, Thr58, or a nonphosphorylated peptide as indicated.
Myc Family Profiling Antibody Sampler Kit: Image 2
Western blot analysis of extracts from various cell lines using c-Myc (E5Q6W) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Myc Family Profiling Antibody Sampler Kit: Image 3
Western blot analysis of extracts from Daudi, EL4, and Raji cells, untreated (-) or treated with MG-132 #2194 (10 μM, 18hr; +), using Phospho-c-Myc (Thr58) (E4Z2K) Rabbit mAb (upper), c-Myc (E5Q6W) Rabbit mAb #18583 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower). Note that Raji cells have a point mutation at Thr58.
Myc Family Profiling Antibody Sampler Kit: Image 4
Western blot analysis of extracts from various cell lines using N-Myc (D4B2Y) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Myc Family Profiling 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.
Myc Family Profiling Antibody Sampler Kit: Image 6
Western blot analysis of various cell lines using L-Myc (E3M5P) Rabbit mAb (upper), c-Myc (E5Q6W) Rabbit mAb #18583 (upper-middle), N-Myc (D4B2Y) Rabbit mAb #51705 (lower-middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The cell type specificity for each of the Myc family members is predicted from RNAseq data and confirms the specificity of the antibodies. NCI-H1963 cells express both wild-type and an RLF-L-Myc fusion protein resulting from intrachromosomal rearrangement.
Myc Family Profiling Antibody Sampler Kit: Image 7
Western blot analysis of extracts from KARPAS-299 and HCT 116 cells, untreated (-) or treated with MG-132 #2194 (10 μM, overnight; +), using Phospho-c-Myc (Ser62) (E1J4K) Rabbit mAb (upper), c-Myc (D84C12) XP® Rabbit mAb #5605 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Myc Family Profiling Antibody Sampler Kit: Image 8
Immunoprecipitation of c-Myc from KG-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is c-Myc (E5Q6W) Rabbit mAb. Western blot was performed using c-Myc (E5Q6W) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Myc Family Profiling Antibody Sampler Kit: Image 9
Western blot analysis of extracts from HDLM-2 cells, untreated (-) or treated with MG-132 #2194 (10 μM, 18 hr; +) or GSK-3 Inhibitor XXII, Compound A (5 μM, 18hr; +), using Phospho-c-Myc (Thr58) (E4Z2K) Rabbit mAb (upper), c-Myc (E5Q6W) Rabbit mAb #18583 (middle) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Myc Family Profiling Antibody Sampler Kit: Image 10
Immunoprecipitation of N-Myc from IMR-32 cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is N-Myc (D4B2Y) Rabbit mAb. Western blot was performed using N-Myc (D4B2Y) Rabbit mAb. A confirmation specific secondary antibody was used to avoid cross reactivity with IgG.
Myc Family Profiling Antibody Sampler Kit: Image 11
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human L-Myc protein (hMYCL-Myc/DDK; +), using L-Myc (E3M5P) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Myc Family Profiling Antibody Sampler Kit: Image 12
Confocal immunofluorescent analysis of SCLC-21H cells (left, positive) and IMR-32 cells (right, negative) using c-Myc (E5Q6W) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
Myc Family Profiling Antibody Sampler Kit: Image 13
Immunohistochemical analysis of paraffin-embedded IMR-32 (left) and HeLa (right) cell pellets using N-Myc (D4B2Y) Rabbit mAb.
Myc Family Profiling Antibody Sampler Kit: Image 14
Immunoprecipitation of L-Myc protein from NCI-H209 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is L-Myc (E3M5P) Rabbit mAb. Western blot analysis was performed using L-Myc (E3M5P) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Myc Family Profiling Antibody Sampler Kit: Image 15
Flow cytometric analysis of IMR-32 cells (blue, negative) and RPMI 8226 cells (green, positive) using c-Myc (E5Q6W) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Myc Family Profiling Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded human neuroblastoma using N-Myc (D4B2Y) Rabbit mAb.
Myc Family Profiling Antibody Sampler Kit: Image 17
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H209 cells and either L-Myc (E3M5P) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human ATF4 promoter primers, human NPM-1 intron 1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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.
Myc Family Profiling Antibody Sampler Kit: Image 18
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT and either c-Myc (E5Q6W) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human ATF-4 Promoter Primers, SimpleChIP® Human NPM1 Intron 1 Primers #4779, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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.
Myc Family Profiling Antibody Sampler Kit: Image 19
Immunohistochemical analysis of paraffin-embedded human testis using N-Myc (D4B2Y) Rabbit mAb.
Myc Family Profiling Antibody Sampler Kit: Image 20
Chromatin immunoprecipitations were performed with cross-linked chromatin from IMR-32 cells and either N-Myc (D4B2Y) Rabbit mAb or N-Myc (D1V2A) Rabbit mAb #84406, 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 MIR17HG, a known target gene of N-Myc (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Myc Family Profiling Antibody Sampler Kit: Image 21
Chromatin immunoprecipitations were performed with cross-linked chromatin from IMR-32 cells and either N-Myc (D4B2Y) Rabbit mAb #51705 or N-Myc (D1V2A) 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 13 (upper), including MIR17HG (lower), a known target gene of N-Myc (see additional figure containing ChIP-qPCR data).
Myc Family Profiling Antibody Sampler Kit: Image 22
Chromatin immunoprecipitations were performed with cross-linked chromatin from IMR-32 cells and either N-Myc (D4B2Y) 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 human MIR17HG intron 1 primers, SimpleChIP® Human MDM2 Intron 2 Primers #90678, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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 # 26717
Cat. # Size Qty. Price
26717T
1 Kit  (6 x 20 microliters) $ 402

Product Description

The Myc Family Profiling Antibody Sampler Kit provides an economical means of detecting Myc family proteins and phosphorylation sites. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Myc Family Profiling Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-c-Myc (Ser62) (E1J4K) Rabbit mAb may not recognize c-Myc phosphorylated at Ser62 when Thr58 is also phosphorylated. Phospho-c-Myc (Thr58) (E4Z2K) Rabbit mAb may also react to c-Myc when dually phosphorylated at Thr58 and Ser62. The phosphorylation site at Thr58 is conserved in N-Myc.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly38 of human N-Myc protein, recombinant proteins specific to the amino terminus of human c-Myc protein and the carboxyl terminus of human L-Myc protein, and synthetic phosphopeptides corresponding to Ser62 and Thr58 of human c-Myc protein.

Background

Members of the Myc/Max/Mad network function as transcriptional regulators with roles in various aspects of cell behavior including proliferation, differentiation, and apoptosis (1). These proteins share a common basic-helix-loop-helix leucine zipper (bHLH-ZIP) motif required for dimerization and DNA-binding. Max was originally discovered based on its ability to associate with c-Myc and found to be required for the ability of Myc to bind DNA and activate transcription (2). Subsequently, Max has been viewed as a central component of the transcriptional network, forming homodimers as well as heterodimers with other members of the Myc and Mad families (1). The association between Max and either Myc or Mad can have opposing effects on transcriptional regulation and cell behavior (1). The Mad family consists of four related proteins; Mad1, Mad2 (Mxi1), Mad3, and Mad4, and the more distantly related members of the bHLH-ZIP family, Mnt and Mga. Like Myc, the Mad proteins are tightly regulated with short half-lives. In general, Mad family members interfere with Myc-mediated processes such as proliferation, transformation, and prevention of apoptosis by inhibiting transcription (3,4).
The Myc family is comprised of c-Myc, N-Myc, and L-Myc with often distinct patterns of expression during development as well as cancer (5). Amplification of each of the family members in cancer is frequently mutually exclusive with c-Myc being the most widely studied and most commonly amplified. N-Myc amplification, on the other hand, is found predominantly in neuroblastomas, and L-Myc amplification has been described in small cell lung cancer (6,7). Phosphorylation of c-Myc at Thr58 and Ser62 can control proteasomal-dependent degradation of the transcription factor. Phosphorylation of c-Myc at these sites is a stepwise process, whereby mitogens, mitosis, or cellular stress induce phosphorylation at Ser62, which serves as a priming site for GSK-3 phosphorylation of Thr58 (8-12).
  1. Baudino, T.A. and Cleveland, J.L. (2001) Mol Cell Biol 21, 691-702.
  2. Blackwood, E.M. and Eisenman, R.N. (1991) Science 251, 1211-7.
  3. Henriksson, M. and Lüscher, B. (1996) Adv Cancer Res 68, 109-82.
  4. Grandori, C. et al. (2000) Annu Rev Cell Dev Biol 16, 653-99.
  5. Nesbit, C.E. et al. (1999) Oncogene 18, 3004-16.
  6. Brodeur, G.M. et al. (1984) Science 224, 1121-4.
  7. Nau, M.M. et al. Nature 318, 69-73.
  8. Lutterbach, B. and Hann, S.R. (1994) Mol Cell Biol 14, 5510-22.
  9. Gregory, M.A. et al. (2003) J Biol Chem 278, 51606-12.
  10. Yada, M. et al. (2004) EMBO J 23, 2116-25.
  11. Seo, H.R. et al. (2008) J Biol Chem 283, 15601-10.
  12. Benassi, B. et al. (2006) Mol Cell 21, 509-19.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

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