Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Knockout Validated, Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western, Chromatin Immunoprecipitation (ChIP)

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP)

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all c-Myc Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human c-Myc
Arg66-Asp201
Accession # P01106

Specificity

Detects human c-Myc in direct ELISAs. Detects human and mouse c-Myc in Western blots. In direct ELISAs, less than 1% cross‑reactivity with recombinant human (rh) L-Myc and rhN-Myc is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse c‑Myc Antibody

Detection of Human c-Myc antibody by Western Blot.

Detection of Human c-Myc by Western Blot.

Western blot shows lysates of LNCaP human prostate cancer cell line and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for c-Myc at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse c-Myc antibody by Western Blot.

Detection of Mouse c‑Myc by Western Blot.

Western blot shows lysates of BaF3 mouse pro-B cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for c-Myc at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of c-Myc-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of c‑Myc-regulated Genes by Chromatin Immunoprecipitation.

HeLa human cervical epithelial carcinoma cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. c-Myc/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thep21promoter was detected by standard PCR.

c-Myc antibody in D3 Mouse Stem Cells by Immunocytochemistry (ICC).

c‑Myc in D3 Mouse Stem Cells.

c-Myc was detected in immersion fixed D3 mouse embryonic stem cell line using Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

c‑Myc antibody in BG01V Human Stem Cells by Immunocytochemistry (ICC).

c‑Myc in BG01V Human Stem Cells.

c-Myc was detected in immersion fixed BG01V human embryonic stem cells using 10 µg/mL Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human c‑Myc antibody by Simple WesternTM.

Detection of Human c‑Myc by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for c-Myc at approximately 78 kDa (as indicated) using 20 µg/mL of Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Shows Human c-Myc Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human c‑Myc Specificity by Using Knockout Cell Line.

Western blot shows lysates of HEK293T human embryonic kidney parental cell line and c-Myc knockout HEK293T cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse c-Myc Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3696) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for c-Myc at approximately 52 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse c-Myc by Western Blot

Detection of Mouse c-Myc by Western Blot

DON significantly inhibits xenograft tumor growth.(A) SK-N-AS tumors were treated with DON at 100 mg/kg or water by i.p. twice weekly. Weight loss in DON-treated mice reduced the treatment cohort to 2 mice indicated by (2) at later timepoints. (B) SK-N-FI tumors were treated with DON at 100 mg/kg, 50 mg/kg or water by i.p. twice weekly or 100mg/kg once a week (100mg/kg; 1x/wk). The * indicates p< 0.05 between control and 50 mg/kg, while ** indicates p<0.05 between control and 100mg/kg; 1x/wk, Student’s t test. (C and D) For SK-N-BE2 and IMR32 subcutaneous xenografts, mice were treated with DON at 50 mg/kg or water control twice weekly by i.p. injections. The indicated tumor cell line was grown to 200 mm3 prior to initiation of treatment and tumor size is given as a percent relative to original tumor volume (% RTV). (E) Western blot analysis of N-Myc and c-Myc expression in a panel of NBL cell lines with beta -tubulin as a loading control. (F) A composite graph of SK-N-FI, SK-N-BE2 and IMR32 tumors treated with DON 50mg/kg or water control twice weekly. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25615615), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse c‑Myc Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: HeLa human cervical epithelial carcinoma cell line chromatin, p21 promoter detected by standard PCR.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed BG01V human embryonic stem cells and D3 mouse embryonic stem cells

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human liver cancer tissue

Knockout Validated

c‑Myc is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in c‑Myc knockout HEK293T cell line.

Simple Western

20 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

0.5 µg/mL
Sample: LNCaP human prostate cancer cell line, HeLa human cervical epithelial carcinoma cell line, and BaF3 mouse pro-B cell line

Reviewed Applications

Read 3 reviews rated 4.7 using AF3696 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: c-Myc

Human c-Myc is a 439 amino acid transcription factor with a bHLH/LZ (basic Helix-Loop-Helix, Leucine Zipper) domain. c-Myc DNA-binding and transcription function is achieved upon heterodimerization with its partner Max. c-Myc is often over-expressed and mutated in hematopoietic tumors. Mutations frequently result in truncations that remove the transactivation region or in the bHLH/LZ domain required for association with Max and DNA. Over the region used as immunogen, human c-Myc is 92% identical to the rat and mouse c-Myc proteins.

Long Name

v-Myc Avian Myelocytomatosis Viral Oncogene Homolog (Avian)

Alternate Names

cMyc, Myc, Myc2, Niard, Nird

Entrez Gene IDs

4609 (Human); 17869 (Mouse); 24577 (Rat)

Gene Symbol

MYC

UniProt

P01106

Additional c-Myc Products

Product Documents for Human/Mouse c‑Myc Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse c‑Myc Antibody

For research use only

Citations for Human/Mouse c‑Myc Antibody

Customer Reviews for Human/Mouse c‑Myc Antibody (3)

4.7 out of 5
3 Customer Ratings
5 Stars
67%
4 Stars
33%
3 Stars
0%
2 Stars
0%
1 Stars
0%

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Human/Mouse c-Myc Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: PC-3 human prostate cancer cell line
    Species: Human
    Verified Customer | Posted 03/01/2019
    Human/Mouse c‑Myc Antibody AF3696
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: U266 and normal T cells
    Species: Human
    Verified Customer | Posted 02/02/2015
    Immunofluorescent staining of multiple myeloma cell line U266 and normal T cells with AF3696
    Human/Mouse c‑Myc Antibody AF3696
  • Name: Anonymous
    Application: Chromatin Immunoprecipitation
    Sample Tested: See PMID 23159369
    Species: Human
    Verified Customer | Posted 01/08/2015

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