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Purified anti-Phosphotyrosine Antibody

Pricing & Availability
Clone
PY20 (See other available formats)
Regulatory Status
RUO
Other Names
p-Tyr
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
1_PY20_Purified_Phosphotyrosine_Antibody_1_050318
Total cell lysate (15 µg protein) from non-treated and 1mM Pervanadate treated HeLa cells (30 min treatment) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with 1:2000 (0.25 µg/mL) diluted purified anti-Phosphotyrosine antibody (clone PY20) (upper). Proteins were visualized using 1:3000 diluted HRP goat anti-mouse-IgG secondary antibody (clone Poly4053, Cat. No. 405306) and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin Antibody (1:2000 diluted, clone 2F1-1, Cat. No. 643807) was used as a loading control (lower). Lane M is the MW ladder.
  • 1_PY20_Purified_Phosphotyrosine_Antibody_1_050318
    Total cell lysate (15 µg protein) from non-treated and 1mM Pervanadate treated HeLa cells (30 min treatment) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with 1:2000 (0.25 µg/mL) diluted purified anti-Phosphotyrosine antibody (clone PY20) (upper). Proteins were visualized using 1:3000 diluted HRP goat anti-mouse-IgG secondary antibody (clone Poly4053, Cat. No. 405306) and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin Antibody (1:2000 diluted, clone 2F1-1, Cat. No. 643807) was used as a loading control (lower). Lane M is the MW ladder.
  • 2_PY20_PURE_Phosphotyrosine_ICC_060118
    NIH3T3 cells were non-treated (A and B) or treated with 1mM pervanadate for 30 minutes (C and D) and then were fixed with 4% paraformaldehyde (PFA) for 10 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained overnight at 4°C with 1: 2500 diluted (0.2 µg/ml, A and C) and 1: 1000 diluted (0.5 µg/ml, B and D) purified anti-Phosphotyrosine antibody, clone PY20 followed by 1:200 diluted (2.5µg/mL) Alexa Fluor® 594 (red) conjugated goat anti-mouse IgG (Cat no. 405326) incubation for one hour at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X.
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309301 25 µg $54
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309302 100 µg $106
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Description

Phosphorylation is a common modification of proteins that can result in alterations in protein function, protein-protein association, cellular localization, and protein-half life. Phosphorylation can occur on threonine, serine, and tyrosine residues. The PY20 monoclonal antibody recognizes phosphorylated tyrosine residues in all species tested (human, mouse, rat, dog, chicken, and frog). The PY20 antibody has been shown to be useful for flow cytometry, immunoprecipitation, Western blotting, and immunofluorescence staining.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat, All Species
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
KLH-conjugated phosphotyrosine
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 50% glycerol. Final antibody concentration is 0.5 mg/ml.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored at -20°C.
Application

WB - Quality tested
ICC - Verified
ICFC, IP- Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. Suggested working dilution(s): Use 0.125 - 1.0 µg per ml antibody dilution buffer per mini-gel. Do not use dilution or blocking buffers containing milk as they may interfere with antibody binding to proteins of interest. Dilution and blocking buffers containing 4% bovine serum albumin are recommended for use with this antibody. For immunocytochemistry, a concentration range of 0.2 - 0.5 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Additional reported applications (for the relevant formats) include: immunoprecipitation1,2, Western blotting1,2, immunocytochemistry3.

Application References

(PubMed link indicates BioLegend citation)
  1. Vuori K, et al. 1995. J. Biol. Chem. 270:22259. (IP, WB)
  2. Glenney J, et al. 1988. J. Immunol. Meth. 109:277. (IP, WB)
  3. Prahalad P, et al. 2004. Am J Physiol Cell Physiol 286:C693. (ICC)
  4. Zentillin L, et al. 2009. FASEB J. 24:1467. PubMed
  5. Philipsen L, et al. 2013. Mol Cell Proteomics. 12:2551. PubMed
  6. Cespedes PF, et al. 2014. PNAS. 111:3214. PubMed
Product Citations
  1. Prochazka R, et al. 2012. Reproduction. 144:535. PubMed
  2. Céspedes P, et al. 2014. Proc Natl Acad Sci U S A. 111:3214. PubMed
  3. Zentilin L, et al. 2009. FASEB J. 24:1467. PubMed
RRID
AB_314763 (BioLegend Cat. No. 309301)
AB_314763 (BioLegend Cat. No. 309302)

Antigen Details

Distribution

Phospho-Specific

Function
Phosphorylation of specific tyrosine residues, signal transduction, cell cycle progression, oncogenic transformation
Biology Area
Cell Biology, Immunology, Signal Transduction
Molecular Family
Phospho-Proteins, Protein Kinases/Phosphatase
Gene ID
5800 View all products for this Gene ID
UniProt
View information about Phosphotyrosine on UniProt.org
Go To Top Version: 2    Revision Date: 03/17/2014

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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