Phospho-c-Myc (T58) Recombinant Rabbit Monoclonal Antibody [SN60-01]
cat.: ET1611-24
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, FC, IHC-P
Clonality: Monoclonal
Clone number: SN60-01
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 49 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Thr58 of Human Myc (P01106-1).
Positive control: HCT 116 cell lysate, HCT 116 treated with 25μM MG-132 for 4 hours cell lysate, NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate, Hela, SKOV-3, human kidney tissue, MCF-7, human skin tissue, mouse skin tissue, rat skin tissue, EL4 cell lysate, EL4 treated with 25μM MG-132 for 4 hours cell lysate.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  FC
  IF-Cell
  IF-Tissue
  IHC-P
1:1,000
1:50-1:100
1:50-1:200
1:50-1:200
1:50-1:500
Uniprot #: SwissProt: P01106 Human | P01108 Mouse | P09416 Rat
Alternative names: Avian myelocytomatosis viral oncogene homolog bHLHe39 c Myc Class E basic helix-loop-helix protein 39 MRTL Myc Myc protein Myc proto oncogene protein Myc proto-oncogene protein myc related translation/localization regulatory factor MYC_HUMAN Myc2 MYCC Niard Nird Proto-oncogene c-Myc Transcription factor p64 v myc avian myelocytomatosis viral oncogene homolog v myc myelocytomatosis viral oncogene homolog
Images
ET1611-24_1.jpg Fig1: Western blot analysis of Phospho-c-Myc (T58) on different lysates with Rabbit anti-Phospho-c-Myc (T58) antibody (ET1611-24) at 1/1,000 dilution.

Lane 1: HCT 116 cell lysate
Lane 2: HCT 116 treated with 25μM MG-132 for 4 hours cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 49 kDa
Observed band size: 57 kDa

Exposure time: 2 minutes 18 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-24) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1611-24_2.jpg Fig2: ICC staining of Phospho-c-Myc (T58) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-24, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1611-24_3.jpg Fig3: ICC staining of Phospho-c-Myc (T58) in SKOV-3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-24, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1611-24_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-c-Myc (T58) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-24, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-24_5.jpg Fig5: Flow cytometric analysis of Phospho-c-Myc (T58) was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-24, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1611-24_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Phospho-c-Myc (T58) antibody (ET1611-24) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-24) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-24_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse skin tissue with Rabbit anti-Phospho-c-Myc (T58) antibody (ET1611-24) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-24) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-24_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat skin tissue with Rabbit anti-Phospho-c-Myc (T58) antibody (ET1611-24) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-24) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1611-24_9.jpg Fig9: Western blot analysis of Phospho-c-Myc (T58) on different lysates with Rabbit anti-Phospho-c-Myc (T58) antibody (ET1611-24) at 1/1,000 dilution.

Lane 1: EL4 cell lysate
Lane 2: EL4 treated with 25μM MG-132 for 4 hours cell lysate
Lane 3: EL4 treated with 25μM MG-132 for 4 hours, then treated with λpp for 1 hour cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 49 kDa
Observed band size: 57 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-24) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.