Anti-Phosphotyrosine antibody [PY20] (ab10321)
![Western blot - Anti-Phosphotyrosine antibody [PY20] (ab10321)](https://www.abcam.com/ps/products/10/ab10321/Images/Phosphotyrosine-Primary-antibodies-ab10321-8.jpg "Western blot - Anti-Phosphotyrosine antibody [PY20] (ab10321)")
Key features and details
- Mouse monoclonal [PY20] to Phosphotyrosine
- Suitable for: WB, ICC/IF, IHC-P, IHC-FoFr
- Reacts with: Species independent
- Isotype: IgG2b
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/IF, IHC-P, IHC-FoFrmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Chemical/ Small Molecule corresponding to Phosphotyrosine conjugated to keyhole limpet haemocyanin.
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General notes
This antibody clone is manufactured by Abcam.
This is a standard clone used to detect phosphotyrosine.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
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... -
Purity
IgG fraction -
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 |
|---|---|---|
| WB | (4) |
Use a concentration of 1 µg/ml.
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| ICC/IF |
1/200.
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| IHC-P |
Use a concentration of 1 µg/ml.
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| IHC-FoFr |
Use a concentration of 1 µg/ml.
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| Notes |
|---|
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WB
Use a concentration of 1 µg/ml. |
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ICC/IF
1/200. |
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IHC-P
Use a concentration of 1 µg/ml. |
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IHC-FoFr
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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Western blot - Anti-Phosphotyrosine antibody [PY20] (ab10321)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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)](https://www.abcam.com/ps/products/10/ab10321/Images/ab10321_1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)This image is courtesy of Sophie Pezet, Univ London Kings Coll, United KingdomImmunofluorescent staining with ab10321 mouse monoclonal [PY20] phosphotyrosine antibody in the rat cortex. Cells stained appear to be microglial cells. Picture taken with objective X20. Protocol: IHC free-floating protocol using 4%PFA fixed brain tissue. Rats were intracardially perfused with 4% PFA. Tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. Primary antibody ab10321 was used at 1ug/ml incubated overnight at room temperature. Secondary antibody was Alexa Fluor 488 used at 1/1000, 2h incubation at room temperature. Image recoloured in Adobe photoshop. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)](https://www.abcam.com/ps/products/10/ab10321/Images/ab10321_2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)This image is courtesy of Sophie Pezet, Univ London Kings Coll, United KingdomImmunofluorescent staining with ab10321 mouse monoclonal [PY20] phosphotyrosine antibody in the rat spinal cord. Cells stained appear to be microglial cells. Picture taken with X40 objective. Protocol: IHC free-floating protocol using 4% PFA fixed spinal cord tissue. Rats were intracardially perfused with 4% PFA. Tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. Primary antibody ab10321 was used at 1ug/ml incubated overnight at room temperature. Secondary antibody was Alexa Fluor 488 used at 1/1000, 2h incubation at room temperature. Image recoloured in Adobe photoshop. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)](https://www.abcam.com/ps/products/10/ab10321/Images/ab10321-17933-anti-phosphotyrosine-antibody-py20-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)IHC image of ab10321 staining in Human Normal Hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10321, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (45)
ab10321 has been referenced in 45 publications.
- Feng X et al. Hypoxia-induced acetylation of PAK1 enhances autophagy and promotes brain tumorigenesis via phosphorylating ATG5. Autophagy N/A:1-20 (2020). PubMed: 32186433
- Chen R et al. CD147 deficiency in T cells prevents thymic involution by inhibiting the EMT process in TECs in the presence of TGFß. Cell Mol Immunol N/A:N/A (2020). PubMed: 31900457
- Lv XX et al. EGFR enhances the stemness and progression of oral cancer through inhibiting autophagic degradation of SOX2. Cancer Med 9:1131-1140 (2020). PubMed: 31823521
- Rimnácová H et al. Low doses of Bisphenol S affect post-translational modifications of sperm proteins in male mice. Reprod Biol Endocrinol 18:56 (2020). PubMed: 32466766
- Yuan G et al. RGS12 is required for the maintenance of mitochondrial function during skeletal development. Cell Discov 6:59 (2020). PubMed: 32922858