Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)
![Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)](https://www.abcam.com/ps/products/185/ab185655/Images/ab185655-349278-AnticMyc-phospho-T58-antibody-EPR17923-dotblot-phospho-peptides.jpg "Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17923] to c-Myc (phospho T58)
- Suitable for: ICC/IF, WB, Flow Cyt, Dot blot
- Reacts with: Human
Overview
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Product name
Anti-c-Myc (phospho T58) antibody [EPR17923]
See all c-Myc primary antibodies -
Description
Rabbit monoclonal [EPR17923] to c-Myc (phospho T58) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate, HeLa cells treated with 200nM Calyculin A and 1uM Okadaic Acid for 60 minutes whole cell lysate. ICC/IF: HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17923 -
Isotype
IgG -
Research areas
Images
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Dot Blot - Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)
Lane 1: c-Myc (phospho T58).
Lane 2: c-Myc (pT58) non-phospho peptide.
Lane 3: c-Myc (pS62) phospho peptide.
Lane 4: c-Myc (pS62) non-phospho peptide.Dot blot analysis using ab185655 at a dilution of 1/1000. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Western blot - Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)](https://www.abcam.com/ps/products/185/ab185655/Images/ab185655-255239-anti-c-myc-phospho-t58-antibody-epr17923-western-blot.jpg)
Western blot - Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)All lanes : Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655) at 1/5000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma ) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma ) treated with 200nM Calyculin A and 1uM Okadaic Acid for 60 minutes whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 49 kDa
Observed band size: 57 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and diluting buffer was 5% NFDM /TBST
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![Western blot - Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)](https://www.abcam.com/ps/products/185/ab185655/Images/ab185655-255240-anti-c-myc-phospho-t58-antibody-epr17923-western-blot.jpg)
Western blot - Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)All lanes : Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with Lambda Phosphatase whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 49 kDa
Observed band size: 57 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and diluting buffer was 5% NFDM /TBST.
The strong band in Lane 1 of WB-2 compared to WB-1 is due to different lysate batches and a lower dilution factor (1:1000).
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![Immunocytochemistry/ Immunofluorescence - Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)](https://www.abcam.com/ps/products/185/ab185655/Images/ab185655-255250-anti-c-myc-phospho-t58-antibody-epr17923-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells (Human epithelial cells from cervix adenocarcinoma) labeling c-Myc (phospho T58) with ab185655 at 1/250, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 (green).
Confocal image showing nuclear staining on HeLa cells. The staining decreased after blocking with phospho peptide (100μg/ml) overnight. The control peptide is a non-phospho peptide.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody -Loading Control (ab7291) at 1/1000 dilution Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/500 (red).
The negative controls are as follows:-
-ve control 1 - ab185655 at 1/500 followed by ab150120 at 1/500.
-ve control 2 -ab7291 at 1/1000 followed by ab150077 at 1/500. -
![Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)](https://www.abcam.com/ps/products/185/ab185655/Images/ab185655-6-benefits-of-recombinant-antibodies.png)
Anti-c-Myc (phospho T58) antibody [EPR17923] (ab185655)
