Anti-Phosphotyrosine antibody [PY20] (ab10321)
Key features and details
- Mouse monoclonal [PY20] to Phosphotyrosine
- Suitable for: WB, ICC/IF
- Reacts with: Species independent
- Isotype: IgG2b
Related conjugates and formulations
Overview
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Product name
Anti-Phosphotyrosine antibody [PY20]
See all Phosphotyrosine primary antibodies -
Description
Mouse monoclonal [PY20] to Phosphotyrosine -
Host species
Mouse -
Tested applications
Suitable for: WB, ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Species independent -
Immunogen
Chemical/ Small Molecule corresponding to Phosphotyrosine conjugated to keyhole limpet haemocyanin.
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Positive control
- ICC/IF: C2C12 cells treated with 2mM H2O2 for 10min. WB: NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Contains 0.4M Arginine -
Concentration information loading...
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Purity
Protein G purified -
Primary antibody notes
This is a standard clone used to detect phosphotyrosine. -
Clonality
Monoclonal -
Clone number
PY20 -
Isotype
IgG2b -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab10321 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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WB | (4) |
Use a concentration of 1 µg/ml.
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ICC/IF |
Use a concentration of 1 µg/ml.
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Notes |
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WB
Use a concentration of 1 µg/ml. |
ICC/IF
Use a concentration of 1 µg/ml. |
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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ab10321 staining Phosphotyrosine in C2C12 cells treated with 2mM H2O2 for 10mins. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab10321 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
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All lanes : Anti-Phosphotyrosine antibody [PY20] (ab10321) at 1 µg/ml
Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate
Lysates/proteins at 5 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Exposure time: 1 minuteCells were serum starved overnight and then incubated at room temperature for 10mins in a final concentration of 1mM sodium vanadate. PDGF was then added at a final concentration of 5ng/ml and cells were incubated at 37ºC for 30mins. Vanadate inhibits endogenous phosphatases and PDGF stimulates phosphorylation. Western blots of NIH 3T3 cell lysates treated with vanadate and PDGF show an array of phosphorylated tyrosine compared to controls.
Protocols
Datasheets and documents
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Datasheet download
References (59)
ab10321 has been referenced in 59 publications.
- Zhu L et al. Myostatin Deficiency Enhances Antioxidant Capacity of Bovine Muscle via the SMAD-AMPK-G6PD Pathway. Oxid Med Cell Longev 2022:3497644 (2022). PubMed: 35663205
- Sano M et al. Neutrophil extracellular traps-mediated Beclin-1 suppression aggravates atherosclerosis by inhibiting macrophage autophagy. Front Cell Dev Biol 10:876147 (2022). PubMed: 35923856
- Allert C et al. Protein tyrosine kinase 2b inhibition reverts niche-associated resistance to tyrosine kinase inhibitors in AML. Leukemia 36:2418-2429 (2022). PubMed: 36056084
- Laudette M et al. Cyclic AMP-binding protein Epac1 acts as a metabolic sensor to promote cardiomyocyte lipotoxicity. Cell Death Dis 12:824 (2021). PubMed: 34471096
- Wang X & Ma Q Wzb of Vibrio vulnificus represents a new group of low-molecular-weight protein tyrosine phosphatases with a unique insertion in the W-loop. J Biol Chem 296:100280 (2021). PubMed: 33450227