EYA3 Polyclonal antibody

EYA3 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human and More (1)

Applications

WB, IHC, IF/ICC, IP, ELISA

Conjugate

Unconjugated

Publications

7

Cat no : 21196-1-AP

Synonyms

EC:3.1.3.48

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Validation Data Gallery

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  WB analysis of HeLa using 21196-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 21196-1-AP (EYA3 Antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

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  WB analysis of L02 using 21196-1-AP

  L02 cells were subjected to SDS PAGE followed by western blot with 21196-1-AP (EYA3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 21196-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 21196-1-AP (EYA3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of L02 using 21196-1-AP

  L02 cells were subjected to SDS PAGE followed by western blot with 21196-1-AP (EYA3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  IP experiment of L02 using 21196-1-AP

  IP result of anti-EYA3 (IP:21196-1-AP, 4ug; Detection:21196-1-AP 1:1000) with L02 cells lysate 1800ug.

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  IHC staining of human breast cancer using 21196-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 21196-1-AP (EYA3 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 21196-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 21196-1-AP (EYA3 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 21196-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 21196-1-AP (EYA3 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 21196-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 21196-1-AP (EYA3 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HeLa using 21196-1-AP

  Immunofluorescent analysis of Hela cells, using EYA3 antibody 21196-1-AP at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red).

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  FC experiment of A431 using 21196-1-AP

  1x10^6 A431 cells were intracellularly stained with 0.25 ug EYA3 Polyclonal antibody (21196-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: HeLa cells, L02 cells
• Positive IP detected in: L02 cells
• Positive IHC detected in: human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: Hela cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:1000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:10-1:100

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 6 publications below
• IHC: See 2 publications below
• IF: See 1 publications below
• IP: See 2 publications below

Product Information

21196-1-AP targets EYA3 in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: EYA3 fusion protein Ag15544
• Full Name: eyes absent homolog 3 (Drosophila)
• Calculated Molecular Weight: 573 aa, 63 kDa
• Observed Molecular Weight: 63-66 kDa
• GenBank Accession Number: BC029500
• Gene Symbol: EYA3
• Gene ID (NCBI): 2140
• RRID: AB_10732957
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

EYA3(Eyes absent homolog 3) belongs to the HAD-like hydrolase superfamily.Its function as histone phosphatase probably explains its role in transcription regulation during organogenesis.EYA3, like EYA1, is expressed in the ninth week of human development and may also underlie developmental defects(PMID:9020840).It has 2 isoforms produced by alternative splicing.This antibody is specific to EYA3.

Publications

Species	Application	Title
human	WB,IHC,IF	Nat Commun; The EYA3 tyrosine phosphatase activity promotes pulmonary vascular remodeling in pulmonary arterial hypertension.; Authors - Yuhua Wang
		Sci Rep; WDR1 is a novel EYA3 substrate and its dephosphorylation induces modifications of the cellular actin cytoskeleton.; Authors - Mihaela Mentel
human	WB,IP	Int J Mol Sci; Analysis of EYA3 Phosphorylation by Src Kinase Identifies Residues Involved in Cell Proliferation.; Authors - Aura E Ionescu
human	WB	PLoS One; Structure-Activity Relationships of Benzbromarone Metabolites and Derivatives as EYA Inhibitory Anti-Angiogenic Agents.; Authors - Ram Naresh Pandey
mouse,human	IP,WB,IHC	Ann Transl Med; Both a hypoxia-inducible EYA3 and a histone acetyltransferase p300 function as coactivators of SIX5 to mediate tumorigenesis and cancer progression; Authors - Chunmei Yang
human	WB	Mol Ther Nucleic Acids; miR-342-5p downstream to Notch enhances arterialization of endothelial cells in response to shear stress by repressing MYC; Authors - Xiaoyan Zhang