Recombinant Anti-Phosphotyrosine antibody [EPR16871] (ab179530)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16871] to Phosphotyrosine
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, ELISA, Dot blot
- Reacts with: Species independent
Related conjugates and formulations
Overview
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Product name
Anti-Phosphotyrosine antibody [EPR16871]
See all Phosphotyrosine primary antibodies -
Description
Rabbit monoclonal [EPR16871] to Phosphotyrosine -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, ELISA, Dot blotmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Whole cell lysate from A431, L6 and NIH/3T3 cells treated with pervanadate. ICC/IF: C2C12 cells treated with Hydrogen peroxide. Flow Cyt (intra): A431 cells treated with pervanadate. IP: Whole cell extract from A431 cells treated with pervanadate.
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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
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16871 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab179530 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/160.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/1000.
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ICC/IF |
1/100.
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IP |
1/100.
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ELISA |
1/6400.
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Dot blot |
1/1000.
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Notes |
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Flow Cyt (Intra)
1/160. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000. |
ICC/IF
1/100. |
IP
1/100. |
ELISA
1/6400. |
Dot blot
1/1000. |
Target
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Relevance
The phosphorylation of specific tyrosine residues has been shown to be a primary mechanism of signal transduction during normal mitogenesis, cell cycle progression and oncogenic transformation, its role in other areas such as differentiation and gap junction communication, is a matter of active and ongoing research. Antibodies that specifically recognize phosphorylated tyrosine residues have proved to be invaluable to the study of tyrosine phosphorylated proteins and the biochemical pathways in which they function.
Images
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (Mouse myoblast cell line) cells labeling Phosphotyrosine with ab179530 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on C2C12 cells is observed. The expression increased after treatment with H2O2 (2mM) for 10 minutes. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:-
-ve control 1: - ab179530 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
Serially diluted ab179530 was bound to immobilised phospho- or control peptides (STAT1 (phospho S727), STAT1 control, STAT5 (phospho T694), STAT5 control); 1 microgram per mL).
The antibody was detected by Goat anti-Rabbit HRPO and signal was developed by TMB substrate.
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Dot blot analysis of INSR/IGF-1R (pY1009) phospho peptide (lane 1) and INSR/IGF-1R non-phospho peptide (lane 2) labelling Phosphotyrosine with ab179530 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Phosphotyrosine was immunoprecipitated from 1mg of A431 (Human epidermoid carcinoma) whole cell extract treated with 1mM pervanadate for 30 minutes with ab179530 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab179530 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: A431 treated with 1mM pervanadate for 30 minutes whole cell extract. Lane 2: PBS instead of A431 whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Multiple bands represent phosph-tyrosine containing proteins precipitated and detected by ab179530. -
All lanes : Anti-Phosphotyrosine antibody [EPR16871] (ab179530) at 1/1000 dilution
Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates treated with 1mM pervanadate for 20minutes
Lane 2 : Untreated Jurkat whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated at 1/1000 dilutionMultiple bands represent phosph-tyrosine containing proteins detected by ab179530
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All lanes : Anti-Phosphotyrosine antibody [EPR16871] (ab179530) at 1/10000 dilution
Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysates treated with 50mM pervanadate for 30 minutes
Lane 2 : Untreated A431 whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilutionMultiple bands represent phosph-tyrosine containing proteins detected by ab179530.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Phosphotyrosine antibody [EPR16871] (ab179530) at 1/1000 dilution
Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates treated with 1mM pervanadate for 10 minutes
Lane 2 : Untreated NIH/3T3 whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilutionMultiple bands represent phosph-tyrosine containing proteins detected by ab179530.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Phosphotyrosine antibody [EPR16871] (ab179530) at 1/1000 dilution
Lane 1 : L6 (Rat skeletal muscle cell line) whole cell lysates treated with 1mM pervanadate for 20 minutes
Lane 2 : Untreated L6 whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilutionMultiple bands represent phosph-tyrosine containing proteins detected by ab179530.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Pervanadate (50mM, 30min.) treated (orange)/untreated (red)A431 (Human epidermoid carcinoma) cells labeling Phosphotyrosine with ab179530 at 1/160 dilutioncompared with a rabbit monoclonal IgG control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
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ELISA analysis of various antigens (1 µg/ml) using ab179530 at 1/6400 dilution followed by Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution.
S/N = signal-to-noise ratio of phospho- versus nonphospho-peptides.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (15)
ab179530 has been referenced in 15 publications.
- Mo X et al. Tyrosine phosphorylation tunes chemical and thermal sensitivity of TRPV2 ion channel. Elife 11:N/A (2022). PubMed: 35686730
- Liu AD et al. Aptamer-SH2 superbinder-based targeted therapy for pancreatic ductal adenocarcinoma. Clin Transl Med 11:e337 (2021). PubMed: 33783993
- Doherty CA et al. Coupled oscillators coordinate collective germline growth. Dev Cell 56:860-870.e8 (2021). PubMed: 33689691
- Barone R et al. Morphological Alterations and Stress Protein Variations in Lung Biopsies Obtained from Autopsies of COVID-19 Subjects. Cells 10:N/A (2021). PubMed: 34831356
- Qiao Y et al. Screening and Functional Analysis of TEK Mutations in Chinese Children With Primary Congenital Glaucoma. Front Genet 12:764509 (2021). PubMed: 34956319