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Neuroscience Neurotransmission Intracellular Signaling Phosphatases
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Anti-Phosphotyrosine antibody [PY20] (ab10321)

  • Datasheet
  • SDS
Reviews (5) Submit a question References (45)

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Western blot - Anti-Phosphotyrosine antibody [PY20] (ab10321)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

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Overview

  • 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.

  • 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

  • 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

    • Neuroscience
    • Neurotransmission
    • Intracellular Signaling
    • Phosphatases
    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Phosphatases
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Amino acid metabolism
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Biotin Anti-Phosphotyrosine antibody [PY20] (ab16388)
    • HRP Anti-Phosphotyrosine antibody [PY20] (ab16389)
    • Alexa Fluor® 647 Anti-Phosphotyrosine antibody [PY20] (ab197508)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)

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.
ICC/IF
1/200.
IHC-P
Use a concentration of 1 µg/ml.
IHC-FoFr
Use a concentration of 1 µg/ml.
Notes
WB
Use a concentration of 1 µg/ml.
ICC/IF
1/200.
IHC-P
Use a concentration of 1 µg/ml.
IHC-FoFr
Use a concentration of 1 µg/ml.

Target

  • 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

  • Western blot - Anti-Phosphotyrosine antibody [PY20] (ab10321)
    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 minute


    Cells 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)This image is courtesy of Sophie Pezet, Univ London Kings Coll, United Kingdom
    Immunofluorescent 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)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phosphotyrosine antibody [PY20] (ab10321)This image is courtesy of Sophie Pezet, Univ London Kings Coll, United Kingdom
    Immunofluorescent 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)
    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

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (45)

Publishing research using ab10321? Please let us know so that we can cite the reference in this datasheet.

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
View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Phosphotyrosine antibody [PY20]

Inconclusive
Abreviews
Abreviews
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain sections)
Specification
Brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Read More

Dr. Sophie Pezet

Verified customer

Submitted Jun 12 2009

Western blot abreview for Anti-Phosphotyrosine antibody [PY20]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (COS1 cells)
Loading amount
10 µg
Specification
COS1 cells
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Apr 07 2009

Western blot abreview for Anti-Phosphotyrosine antibody [PY20]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293)
Loading amount
10 µg
Specification
HEK 293
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Apr 07 2009

Western blot abreview for Anti-Phosphotyrosine antibody [PY20]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Testis)
Loading amount
200 µg
Specification
Testis
Gel Running Conditions
Reduced Denaturing (10% acrylamide)
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Oct 28 2008

Western blot abreview for Anti-Phosphotyrosine antibody [PY20]

Below Average
Abreviews
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (Human Aortic Endothelial Cell)
Loading amount
30 µg
Specification
Human Aortic Endothelial Cell
Treatment
200ng/ml VEGF
Gel Running Conditions
Reduced Denaturing (4-20% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Aug 08 2008

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